Toward stomatal-flux based forest protection against ozone: The MOTTLES approach.

European standards for the protection of forests from ozone (O3) are based on atmospheric exposure (AOT40) that is not always representative of O3 effects since it is not a proxy of gas uptake through stomata (stomatal flux). MOTTLES "MOnitoring ozone injury for seTTing new critical LEvelS" is a LIFE project aimed at establishing a permanent network of forest sites based on active O3 monitoring at remote areas at high and medium risk of O3 injury, in order to define new standards based on stomatal flux, i.e. PODY (Phytotoxic Ozone Dose above a threshold Y of uptake). Based on the first year of data collected at MOTTLES sites, we describe the MOTTLES monitoring station, together with protocols and metric calculation methods. AOT40 and PODY, computed with different methods, are then compared and correlated with forest-health indicators (radial growth, crown defoliation, visible foliar O3 injury). For the year 2017, the average AOT40 calculated according to the European Directive was even 5 times (on average 1.7 times) the European legislative standard for the protection of forests. When the metrics were calculated according to the European protocols (EU Directive 2008/50/EC or Modelling and Mapping Manual LTRAP Convention), the values were well correlated to those obtained on the basis of the real duration of the growing season (i.e. MOTTLES method) and were thus representative of the actual exposure/flux. AOT40 showed opposite direction relative to PODY. Visible foliar O3 injury appeared as the best forest-health indicator for O3 under field conditions and was more frequently detected at forest edge than inside the forest. The present work may help the set-up of further long-term forest monitoring sites dedicated to O3 assessment in forests, especially because flux-based assessments are recommended as part of monitoring air pollution impacts on ecosystems in the revised EU National Emissions Ceilings Directive.

A new completely flexible ring for mitral valve annuloplasty: the "Rama-Valvuloplasty-Ring". Five-year Pitié Salpétrière experience.

The aim of this study was to analyze the effects about the use of a new completely flexible ring for mitral valve anuloplasty, the "Rama-Valvuloplasty-ring", in 182 patients operated on in the Pitié-Salpétrière Hospital, Paris, France for mitral valve regurgitation (MVR). From January 1998 to December 2003, 182 patients with mitral regurgitation (MR) underwent mitral reconstructive surgery at our institution with the "Rama-Valvuloplasty-Ring". This group was made up of 117 men (64.3%) and 65 women (35.7%). The age ranged from 19 to 87 years (mean 62.51 +/- 8.2 years). The patients surviving the operation were the subject of a prospective follow-up. In the preoperative stage sinus rhythm was found in 71.97% (131) of patients and atrial fibrillation in the remaining 28.03% (51) of patients. The mean NYHA FC was 2.9 +/- 1.7 and subdivided as follows: 65 patients in FC I-II (35.72%), 104 patients in FC III (57.14%) and 13 patients in FC IV (7.14%). Most of the patients have shown, in the preoperative echocardiogram, grade II M.R. (46.15% N. 84) and grade III M.R. (29.12%, N.53); 24.72% of the patients (N. 45) had grade IV M.R. The mean E.F. was 42.8 +/- 9.7%. Left ventricular end diastolic diameter (LVEDD) was 57.7 +/- 9.7 mm. The causes of mitral valve insufficiency were degenerative disease in 141 patients (77.47%), post-ischemic disease in 21 patients (11.53%), rheumatic valvular disease in 11 patients (6.05%) and infectious endocarditis in 9 patients (4.95%). All the patients were operated using the Rama-Valvuloplasty-Ring. Ring sizes most commonly used were 30 mm and 32 mm, respectively in 92 patients (50.55%) and 41 patients (22.54%), followed by 28 mm (43 patients, 23.62%), 34 mm (5 patients, 2.74%), 36 mm (1 patient, 0.55%). The surgical tecnique was valve quadrangular resection in 103 patients (56.60%), triangular resection in 57 patients (31.32%) and no valve resection in 22 patients (12.08%). Among the above, 89 patients (48.90%) underwent an associated intervention as follows: 44 patients (24.18%) underwent coronary revascularization: 18 patients (9.89%) with single by-pass surgery, 21 patients (11.54%) with double by-pass, 5 patients with triple by-pass (2.75%); 42 patients (23.07) underwent aortic valve replacement (AVR); 3 patients (1.65%) underwent aortic repair. Early postoperative mortality was 2.19% (4 of 182 patients). Early postoperative echocardiographic control showed MR grade 0 in 142 patients (79.78%) and grade I in 36 (20.22%) with mean grade 0.4 +/- 0.12; no patients with grade III or IV. Therefore, there was no mitral annuloplasty failure requiring valve replacement (MVR). During the follow-up there were 12 late deaths (12 of 178 patients, 6.74%). Only one death was valve-related (thrombosis) whereas the other 11 ones were non cardiac-related deaths (subdural frontal haematoma, septic shock). Postoperative transthoracic echocardiogram data were available in 166 patients at 5 years: the presence of postoperative MR was evaluated and severity was graded as mild in 33 patients (19.88%), moderate in 18 patients (10.84%), severe in 3 (1.81%) patients. There was no MR in the other 112 patients (67.47%); LVEDD was 49.4 +/- 6.5 mm; EF was 51.8 +/- 4.3%. The mean NYHA FC was 0.8 +/- 0.4. Only one patient was reoperated on during the follow-up because of mitral annuloplasty failure with MVR. In conclusion, mid-term 5-years follow up is good for patients operated on with the new completely flexible Rama-Valvuloplasty-Ring for mitral annuloplasty. This study has also verified the advantage about the Rama-Valvuloplasty-Ring use in the preservation of native valve apparatus.

The endovascular treatment of traumatic isthmic aortic rupture: three years follow-up.

The isthmic aortic rupture represents the main cause of death in car crash accidents, because of closed chest trauma. Early medical and surgical care and endovascular prosthesis treatment with semi-invasive method can improve short and mid term survival. Nine patients with traumatic isthmic aortic rupture underwent endoprosthesis aortic implantation. All the patients were male, mean age 42.48 +/- 17.66 years. Operations included 5 acute cases and 4 chronic cases (chance diagnosis). In all cases the diagnosis was performed by tomodensitometric exam. Cloth prostheses were used (self-expansible Goretex- or Dacron-stent). Three years after the endoprosthesis implantation, we obtained the complete thrombosis of the false aortic lumen in all patients, both acute and chronic, as well as the levelling of the false aneurysms without complications of any kind.

Interferon-free therapy in hepatitis C virus mixed cryoglobulinaemia: a prospective, controlled, clinical and quality of life analysis.

BACKGROUND: Cryoglobulinaemic vasculitis (CV) is a lymphoproliferative disorder related to hepatitis C virus (HCV) infection; anti-viral therapy is the first therapeutic option. CV can be incapacitating, compromising the patients' quality of life (QoL). In a controlled study, interferon-based therapy was associated with a lower virological response in vasculitic patients than in patients without vasculitis. Limited, uncontrolled data on direct-acting anti-virals are available. AIM: To evaluate safety, clinical efficacy, virological response and the impact of interferon-free treatment on QoL in HCV patients with and without mixed cryoglobulinaemia (MC). METHODS: We prospectively studied HCV patients with cryoglobulinaemia (with vasculitis-CV- and without vasculitis-MC-) and without cryoglobulinaemia (controls), treated with direct-acting anti-virals. Hepato-virological parameters, CV clinical response and impact on QoL were assessed. RESULTS: One hundred and eighty-two HCV patients were recruited (85 with CV, 54 with MC and 43 controls). A sustained virological response at 12 weeks (SVR12) was achieved in 166 (91.2%) patients (77/85 CV, 48/54 MC, 41/43 controls). In CV SVR patients, cryocrit levels progressively decreased and clinical response progressively improved, reaching 96.7%, 24 weeks after treatment. QoL, baseline physical and mental component summaries were lower in the CV group compared to the other groups (P < 0.05). Scores improved in all groups, and significantly in CV patients after SVR. CONCLUSIONS: No significant differences in SVR rates were recorded between cryoglobulinaemic patients and controls and a high clinical and immunological efficacy was confirmed in CV, supporting the role of interferon-free therapy as the first therapeutic option. Interestingly, CV patients had worse baseline QoL than other HCV-positive groups and interferon-free therapy was effective in significantly increasing QoL, suggesting the important role of direct-acting anti-viral-based therapy in improving CV's individual and social burden.

Placental neurokinin B mRNA expression increases at preterm labor.

INTRODUCTION: Neurokinin B (NKB) is a neuropeptide belonging to the family of tachykinins-related peptides that elicits contractility of human myometrial strips in vitro. The present study evaluates whether placental mRNA and peptide expression of NKB change in women at preterm labor. METHODS: A group of 26 women with singleton pregnancies were enrolled in the study. Placental tissue specimens were collected from pregnant women delivering after elective cesarean section, after labor at term, or after preterm labor. Changes in placental NKB mRNA and protein expression were evaluated by real-time quantitative RT-PCR analysis and by immunofluorescence respectively. RESULTS: Placental mRNA expression of NKB was significantly higher after term and preterm labor (P<0.001) than cesarean section, and highest after preterm labor. Immunofluorescent staining in placentas from preterm or term labor was more intense than after cesarean section (P<0.001). In particular, NKB protein expression was higher in placentas collected after preterm labor than those collected after term labor. DISCUSSION: Neurokinin B mRNA and protein are highly expressed in placenta at term and preterm labor; thus, the involvement of this neuropeptide in the events cascade leading to parturition may be suggested.

Maternal plasma corticotropin-releasing factor (CRF) and CRF-binding protein (CRF-BP) levels in post-term pregnancy: effect of prostaglandin administration.

OBJECTIVE: Placental corticotropin-releasing factor (CRF) affects myometrial contractility and the secretion of several uterotonins such as prostaglandins (PGs); however, the activity of CRF is counteracted by CRF-binding protein (CRF-BP). At term and pre-term labor, CRF levels in maternal plasma are highest whereas those of CRF-BP are falling, and the cause of this fall is unknown. Thus, in this study, we investigated the effect of PG administration for labor induction on maternal plasma CRF and CRF-BP concentrations. DESIGN: Maternal plasma CRF and CRF-BP levels were assayed before and after (2 h later) induction of labor by intracervical administration of prostaglandin E(2) (PGE(2)), and at delivery in a group of healthy post-term pregnancies (n=18). Controls were women at term out of labor (n=22), who subsequently progressed to deliver a healthy singleton baby. METHODS: CRF was measured by two-site immunoradiometric assay, and CRF-BP was assayed by radioimmunoassay. RESULTS: Maternal plasma CRF levels were significantly (P<0.0001) lower and CRF-BP significantly (P<0.0005) higher in post-term than in term pregnancies. With respect to induction of labor, 2 mg PGE(2) were sufficient to increase maternal plasma CRF levels at delivery (P<0.005). While 0.5 mg PGE(2) significantly decreased maternal plasma CRF-BP levels at delivery (P<0.001), 2.0 mg PGE(2) significantly reduced CRF-BP concentrations both after 2 h (P<0.05) and at delivery (P<0.0001). CONCLUSIONS: In the light of the well-known stimulation of prostaglandin release by CRF, these data suggest a positive feedback effect of PGE(2) on maternal CRF release during induced labor.

Changes in Fe site structure from fetal to adult hemoglobin probed by XANES.

Iron X-ray absorption near edge structure (XANES) spectra of human fetal (F) and adult (A) deoxyhemoglobin (deoxyHb) measured at the Frascati synchrotron radiation facility reveal the different geometrical structure of the Fe-porphyrin complexes in the two proteins. By this method, having determined for the first time the variation of atomic positions in fetal and adult hemoglobin in solution (close to the 'in vivo' situation), we give further insight into the structure-function relationship in hemoglobins.

Heterogeneity of the isolated subunits of the fetal and adult human hemoglobin in solution, detected by XANES spectroscopy.

Differences in the local structure of the heme in the isolated alpha-, beta- and gamma-chains of the adult and fetal human hemoglobin are detected by XANES (X-ray absorption near-edge structure) spectroscopy. The ligand bonding angle to the iron ion in the ligated forms and the displacement of the Fe respect to the porphyrin plane in the deoxy forms are found to be different for each chain.

X-ray absorption near edge structure (XANES) determination of calcium sites of troponin C and parvalbumin.

Using synchrotron radiation at the Frascati storage ring ADONE, the X-ray Absorption Near Edge Structure (XANES) has been applied to determine homologies and modifications of the local structure of the calcium binding sites of troponin C. In all four calcium binding sites, Ca2+ appears to be co-ordinated to carboxyl and carbonyl groups in a characteristic configuration. No structural difference has been found between high and low-affinity sites. A distortion of the Ca2+ site geometry by binding of Mg2+ has been observed. The XANES of parvalbumin has been measured and found to be different from troponin C. A tentative identification of the characteristic XANES spectra of the two different Ca2+ sites in this protein is reported.

Regulation of acetylcholine receptor desensitization in mouse myotubes by cytosolic cyclic AMP.

Whole-cell currents activated by bath applications of acetylcholine (ACh) (10-30 microM) were recorded from patch-clamped myotubes of the mouse C2 cell line. Increasing concentrations of forskolin caused a dose-dependent fast decay of ACh-activated currents as compared to the long-lasting ACh-currents in control cells. The forskolin-induced modulation of nicotinic ACh receptor (nAChR) desensitization was proportional to the drug-induced elevation in the cyclic AMP (cAMP) cellular content. Furthermore, an increase in the rate of decay of the ACh-current response, which paralleled an elevation in cAMP cellular content, was caused by treatment with a calcitonin gene-related peptide (1 microM), 8-Br-cAMP (0.5 mM), or by loading the myotubes with cAMP. These results therefore indicate that the desensitization of nAChR is a cAMP-related process in C2-myotubes.

Energy metabolism of human LoVo colon carcinoma cells: correlation to drug resistance and influence of lonidamine.

The relationship between modification of energy metabolism and extent of drug resistance was investigated in two sublines (LoVoDX and LoVoDX10) from human LoVo colon carcinoma cells that exhibit different degrees of resistance to doxorubicin. Results indicated that the extent of alteration in energy metabolism strictly correlated with degree of resistance. In LoVoDX cells, only 14CO2 production was enhanced, whereas in the more resistant LoVoDX10 cells, both 14CO2 and aerobic lactate production were stimulated. The basal and glucose-supported efflux rate and the amount of drug extruded by LoVoDX10 cells were significantly higher than in the resistant LoVoDX cells. Because the expression of surface P-170 glycoprotein was similar in both cell lines, this phenomenon was attributed to increased efflux pump activity resulting from greater ATP availability. Inhibition of 14CO2 production, aerobic glycolysis, and clonogenic activity by lonidamine (LND) increased with enhancement of the energy metabolism. Moreover, LND, by affecting energy-yielding processes, reduced intracellular ATP content, lowered the energy supply to the ATP-driven efflux pump, and inhibited, almost completely, doxorubicin extrusion by resistant LoVo cells. These findings strongly suggest that LND, currently used in tumor therapy, reduces drug resistance by restoring the capacity to accumulate and retain drug of cells with the MDR phenotype that overexpress P-170.

Effects of long-term ambient ozone exposure on biomass and wood traits in poplar treated with ethylenediurea (EDU).

This is the longest continuous experiment where ethylenediurea (EDU) was used to protect plants from ozone (O3). Effects of long-term ambient O3 exposure (23 ppm h AOT40) on biomass of an O3 sensitive poplar clone (Oxford) were examined after six years from in-ground planting. Trees were irrigated with either water or 450 ppm EDU. Above (-51%) and below-ground biomass (-47%) was reduced by O3 although the effect was significant only for stem and coarse roots. Ambient O3 decreased diameter of the lower stem, and increased moisture content along the stem of not-protected plants (+16%). No other change in the physical wood structure was observed. A comparison with a previous assessment in the same experiment suggested that O3 effects on biomass partitioning to above-ground organs depend on the tree ontogenetic stage. The root/shoot ratios did not change, suggesting that previous short-term observations of reduced allocation to tree roots may be overestimated.

Acetylcholine-activated inward current induces cytosolic Ca2+ mobilization in mouse C2C12 myotubes.

We examined the spatiotemporal pattern of intracellular Ca2+ liberation in mouse myotubes by means of fluorescence imaging of cytosolic free Ca2+ together with the simultaneous recording of membrane whole-cell currents. Acetylcholine (ACh) applications to C2C12 myotubes equilibrated in Ca(2+)-free medium and voltage clamped at -50 mV evoked localized fluorescence transients of variable amplitude with less than 0.5 s delay. Under the same experimental conditions, fluorescence transients were elicited by ACh also in mouse primary myotubes. Ca2+ transients were inhibited in myotubes clamped at depolarized potentials (-10 mV to +50 mV), or equilibrated in a Na+,Ca(2+)-free medium as well as in cells loaded with heparin, or with inositol (1,4,5) trisphosphate (InsP3). To investigate whether InsP3 could induce Ca2+ mobilization, [Ca2+]i determinations were carried out in myotubes loaded with InsP3 through the whole-cell patch-clamp recording pipette or by extracellular application in permeabilized cells. InsP3 diffusion into the myoplasm caused Ca2+ spikes with 5 +/- 1 s (mean +/- SEM) delay from the rupture of the membrane patch. Spikes were followed by sustained increases in fluorescence or by damped oscillations. In permeabilized myotubes, InsP3 induced the release of sequestered 45Ca2+ with a half-maximally effective concentration (EC50) of 0.28 +/- 0.05 microM, and Hill coefficient of 0.79 +/- 0.09. It is concluded that the ACh-activated inward current in mouse myotubes is coupled to cytosolic Ca2+ mobilization from internal InsP3-sensitive pools.

XANES study of iron displacement in the haem of myoglobin.

The XANES (X-ray absorption near edge structure) spectra of deoxy human adult haemoglobin (HbA) and myoglobin (Mb) have been measured at the wiggler beam line of the Frascati synchrotron radiation facility. The XANES are interpreted by the multiple scattering cluster theory. The variations in the XANES between HbA and Mb are assigned to changes in the Fe-porphyrin geometry.

CXC chemokines interleukin-8 (IL-8) and growth-related gene product alpha (GROalpha) modulate Purkinje neuron activity in mouse cerebellum.

We give here evidence that Purkinje neurons (PNs) of mouse cerebellar slices studied with patch clamp technique combined with laser confocal microscopy, respond to human IL-8 and GROalpha by (i) a cytosolic Ca2+ transient compatible with inositol (1,4,5) trisphosphate (InsP3) formation; (ii) an enhancement of the neurotransmitter release; and (iii) an impairment of the long-term depression of synaptic strength (LTD). It was also found the expression of IL-8 receptor type 2 in PN and granule cells by immunofluorescence, immunoblotting and RT-PCR analysis. Considered together these findings suggest that IL-8 and GROalpha may play a neuromodulatory role on mouse cerebellum.

Platelet activation and cytokine production during hypothermic cardiopulmonary bypass--a possible correlation?

Cardiopulmonary bypass (CPB) is associated with impaired platelet function and a systemic inflammatory response. The present study was designed to evaluate whether any correlation between platelet activation and inflammatory response during CPB exists. The results obtained from 8 patients undergoing hypothermic CPB for cardiac surgery showed the occurrence of a moderate degree of platelet activation during CPB, demonstrated by an increase of platelet CD62P expression in correlation with an increase of beta-thromboglobulin levels, with a concomitant decrease of in vitro platelet response. Plasma IL-1beta levels significantly increased during CPB, with a peak between 1 and 4 h after CPB. Similarly, IL-6 levels were elevated 30 min from CPB starting, peaked at 4 h, and remained elevated after 24 h. A direct correlation was found between plasma IL-1beta and IL-6 levels. A significant correlation between plasma IL-1beta and beta-thromboglobulin levels was also found. In turn, plasma beta-thromboglobulin levels correlated with CD62P expression on activated platelets. An inverse correlation was found between in vitro platelet aggregation and plasma IL-1beta or IL-6 levels. From the present results it may be speculated that platelet activation during CPB may contribute, through the release of IL-1beta, to activation of endothelial cells and subsequent release of other cytokines with chemotactic and pro-inflammatory properties, thus playing an important role in the inflammatory response associated with CPB.

Modulation of the neurotransmitter release in rat cerebellar neurons by GRO beta.

We report here that, in cultured cerebellar granule cells, the CXC chemokine GRObeta stimulates the signaling pathway of the extracellular signal-regulated kinases, and enhances both evoked and spontaneous postsynaptic currents in patch clamped Purkinje neurons from rat cerebellar slices. The GRObeta-induced enhancement of the excitatory post-synaptic currents evoked by stimulating the parallel fibres is blocked by the inhibitor of the extracellular signal-regulated kinases pathway PD98059, which also reduces both basal frequency of spontaneous post-synaptic currents and mean amplitude of evoked excitatory post-synaptic currents. Our results suggest that GRObeta modulates neurotransmitter release in the cerebellum through the activation of the extracellular signal-regulated kinases pathway.

[Analysis of risk factors in patients undergoing closed heart mitral commissurotomy]. / Analisi dei fattori di rischio in pazienti sottoposti a commissurotomia mitralica a cuore chiuso.

The aim of this study was to analyse the risk factors in patients undergoing closed heart commissurotomy (CHC). From 1956 to 1978 a total of 630 patients with severe mitral stenosis underwent CHC at this Institute. The study was performed on a sample of 100 patients. Follow-up was performed using data taken from the Institute archives, questionnaires sent to the doctors in charge, telephone interviews or visits to outpatient clinics by the Institute's medical staff. The mortality rate 30 days after surgery was 2.9%. Re-CHC was necessary in 11 patients after a mean interval of 7.2 years. The overall probability of survival was 83%, 71%, 48% an 33% respectively at 10, 20, 30 and 40 years. Survival free from mitral re-operation at 10, 20, 30 and 40 years was 76%, 39%, 29% and 11% respectively. The probability of incidence for central and peripheral thromboembolism was 38%, 26%, 11% and 4% at 10, 20, 30 and 40 years. Mitral valve replacement surgery (MVR) was performed in a total of 64 patients. Operative mortality was 0.6% (1 patient). The mean duration of follow-up in patients undergoing post-CHC MVR was 10 years. The probability os survival in overall terms and free of cardiac decompensation was 79 and 53% respectively. From this study it is clear the CHC is a surgical procedure that offers excellent long-term results with a low incidence od thromboembolic events, very low costs and a good quality of life. CHC currently represents a valid alternative o mitral valve replacement in selected patients.

[Primary malignant lymphoma of the parotid gland. Report of a case]. / I linfomi maligni primitivi della parotide. A proposito di un caso.

Primary malignant lymphoma of the parotid gland is often correlated, in literature data, with inflammatory autoimmunological disease of the salivary gland. Finding of blasticlymphoid cells in myoepithelial scialoadenitic areas in a case of parotid disease is described by the Authors. Needle may be useful in detecting these high radio-chemosensitive neoplasms to reduce the occurrence of complications of total parotidectomy.

Corticotrophin-releasing factor and urocortin inhibit system A activity in term human placental villous explants.

Plasma corticotrophin-releasing factor (CRF) and urocortin are elevated in preterm labour and/or fetal growth restriction (FGR). FGR is associated with reduced placental system A amino acid transporter activity and in vitro data suggest altered endocrine status could be responsible. Here we test the hypothesis that CRF and urocortin inhibit placental system A activity. Chronic (48h) exposure of term placental villous explants to these hormones (10(-7)M) significantly reduced system A activity (Na(+)-dependent (14)C-methylaminoisobutyric acid uptake), whereas 1h exposure had no effect. We propose elevated CRF and urocortin contribute to FGR through negative regulation of placental system A activity.