Instrumental monitoring of the birth and development of truffles in a Tuber melanosporum orchard.

Mycorrhizal symbiotic plants, soil suitability, temperature, and humidity are, by general consensus, considered decisive factors in truffle production. However, experimental approaches to define the environmental conditions that stimulate formation of truffle primordia and promote their growth to maturity have been lacking. By analysis of data of many atmospheric and soil parameters collected since 2009 within a Tuber melanosporum orchard, the trends of metabolic activity, detected as CO2 production in the soil, have been identified as the most reliable parameter to indicate the 'birth' of the truffle primordia. They seem to be produced when mycelial activity is intense and undergoes water stress, after which it resumes. About 6-18 days after recovery of metabolic activity, we could collect primordia of T. melanosporum. Many die or develop too early and consequently rot or are eaten by insect larvae. These events occur several times during summer and autumn, those that 'sprout' in late summer or later grow steadily and reach maturity. Using a particular ground-penetrating radar (GPR) setup to discriminate truffles, we could identify individual truffles in the soil after they have enlarged to at least 6 mm in diameter and follow their growth in volume and diameter over time. These two instrumental methods (CO2 sensor and GPR), although yet to be improved, open new important perspectives to better understand truffle biology and manage truffle orchards to support the newly acquired demonstration of the fundamental role of host plants for the nutrient transfer to the ectomycorrhiza-mycelium-fruiting body complex of T. melanosporum.

Metabonomic analysis of exhaled breath condensate in adults by nuclear magnetic resonance spectroscopy.

Exhaled breath condensate (EBC) is a noninvasive method for the study of airway lining fluid. Nuclear magnetic resonance (NMR) spectroscopy can provide biochemical profiles of metabolites in biological samples. The aim of the present study was to validate the NMR metabonomic analysis of EBC in adults, assessing the role of pre-analytical variables (saliva and disinfectant contamination) and the potential clinical feasibility. In total, 36 paired EBC and saliva samples, obtained from healthy subjects, laryngectomised patients and chronic obstructive pulmonary disease patients, were analysed by means of (1)H-NMR spectroscopy followed by principal component analysis. The effect on EBC of disinfectant, used for reusable parts of the condenser, was assessed after different washing procedures. To evaluate intra-day repeatability, eight subjects were asked to collect EBC and saliva twice within the same day. All NMR saliva spectra were significantly different from corresponding EBC samples. EBC taken from condensers washed with recommended procedures invariably showed spectra perturbed by disinfectant. Each EBC sample clustered with corresponding samples of the same group, while presenting intergroup qualitative and quantitative signal differences (94% of the total variance within the data). In conclusion, the nuclear magnetic resonance metabonomic approach could identify the metabolic fingerprint of exhaled breath condensate in different clinical sets of data. Moreover, metabonomics of exhaled breath condensate in adults can discriminate potential perturbations induced by pre-analytical variables.

Validation study of nasal nitric oxide measurements using a hand-held electrochemical analyser.

BACKGROUND: Exhaled nitric oxide (NO) measurement is a simple and non-invasive method for monitoring airway inflammation. Similarly, nasal NO has been proposed as a surrogate marker in inflammatory diseases of the upper airways, e.g. allergic rhinitis. A new portable analyser using an electrochemical sensor has been developed for measurements of exhaled NO, and its reproducibility and comparison with other analysers has been tested recently in healthy subjects and in patients with lower airways disease. The application of this hand-held analyser in nasal NO analysis was tested and compared to the gold standard represented by a chemiluminescence analyser. MATERIALS AND METHODS: Thirty subjects including 15 patients with allergic rhinitis (AR) and 15 healthy subjects (HS) were studied. The intraindividual variability, calculated as the difference in nasal NO levels between two measurements from a single nasally exhaled breath manoeuvre, and the comparison between the electrochemical analyser (NIOX MINO, Aerocrine) and a chemiluminescence analyser (NOA, Sievers) were performed. RESULTS: In AR patients mean nasal NO was 59.0 +/- 16.3 p.p.b. with the MINO and 58.3 +/- 15.6 p.p.b. with the NOA. In HS nasal NO was 49.1 +/- 10.8 p.p.b. with the MINO and 49.8 +/- 8.2 p.p.b. with the NOA. The Bland-Altman analysis showed bias values of 0.005 +/- 3.6 with the 95% limits of agreement from -6.97 to 6.98 p.p.b. CONCLUSION: Measurements of nasal NO levels with a hand-held electrochemical analyser are reproducible and the results are comparable to a stationary chemiluminescence analyser.

CD8+ T-cell alveolitis in familial pulmonary alveolar microlithiasis.

Pulmonary alveolar microlithiasis (PAM) is a rare diffuse lung disease characterised by the accumulation of calcium phosphate microliths within the alveoli. The causative mechanism of PAM has only recently been discovered, and involves a gene mutation of sodium phosphate co-transporter, which is expressed by alveolar epithelial cells. This mutation may have variable consequences on the clinical phenotype. However, pulmonary cell immune phenotyping in familial PAM has not previously been assessed. In the present article, the analysis of bronchoalveolar lavage fluid of two siblings with PAM diagnosis revealed a pattern of lymphocytic alveolitis with accumulation of CD8+ T-cells. The clonal complexity of this lymphocyte's population was assayed by spectratyping, which showed an oligoclonal accumulation of T-cells with a restricted variable beta T-cell receptor (TCR) gene usage. TCR analysis in peripheral blood lymphocytes revealed no abnormal patterns of T-lymphocytes. In the pulmonary alveolar microlithiasis familial cases reported, CD8-mediated maladaptive immune response may have taken place in the bronchoalveolar compartment. The relationship between this immune dysregulation and genetic background in pulmonary alveolar microlithiasis warrants further investigation.

Exhaled breath condensate as matrix for toluene detection: a preliminary study.

The study was designed to investigate whether exhaled breath condensate, obtained by cooling exhaled air in spontaneous breathing, could be a suitable matrix for toluene quantitative analyses. Nine healthy subjects were exposed for a short period (20 min) to a known concentration of toluene. Exhaled breath condensate samples were collected before and at the end of the exposure, while the environmental concentration of toluene was continuously monitored. Toluene was analysed by head-space gas-chromatography mass spectrometry, and assay repeatability was also estimated in vitro. Baseline and post-exposure measurement of hippuric acid, the urinary toluene metabolite, was performed to assess current toluene exposure. Before the exposure toluene concentrations in the exhaled breath condensate were lower than the detectable limit in all subjects, while after the exposure toluene was detectable with a median value 0.35 microg l-1 (range 0.15-0.55 microg l-1) in all the exhaled breath condensate samples. As compared with the standard calibration in distilled water, the curves obtained by exhaled breath condensate were linear and comparable with the range examined in vivo for toluene. A significant correlation was found between the environmental toluene levels and toluene in the exhaled breath condensate at the end of exposure. Furthermore, a significant relationship between increased exhaled breath condensate toluene levels and urinary hippuric acid after the exposure was found. In conclusion, exhaled breath condensate is a promising matrix for toluene assessment, although its application in humans requires further investigations.

Humming-induced release of nasal nitric oxide for assessment of sinus obstruction in allergic rhinitis: pilot study.

BACKGROUND: Humming greatly increases nasal nitric oxide (NO) in healthy people by causing a rapid washout of NO from the sinuses. This increase is abolished in patients with complete sinus ostial obstruction. OBJECTIVE: Allergic rhinitis is a risk factor for development of sinusitis and we wanted to study whether nasal NO measurement during humming could be used to detect sinus abnormalities in this disorder. METHODS: Fifty-nine consecutive subjects with mild to moderate allergic rhinitis were studied. Their present nasal symptoms were recorded. Then NO levels were measured by chemiluminescence during quiet single-breath nasal exhalations and humming exhalations at a fixed exhalation flow of 0.2 L s(-1). Based on the NO results the patients were divided into two groups: those with a great increase in nasal NO during humming (humming responders, n = 46) and those without a significant increase (humming nonresponders, n = 13). In 11 of the nonresponders and in 22 of the responders the passage to the osteomeatal complex area was assessed and scored by nasal endoscopy. This was carried out by an oto-rhino-laryngologist unaware of the NO results. RESULTS: Among the nonresponders nine of 11 patients (80%) had endoscopic signs of bilateral sinus obstruction, compared with one of the 22 (< 5%) humming responders. Baseline nasal symptom score and NO levels during quiet exhalation were not significantly different between the groups CONCLUSION: Absence of a nasal NO peak during humming is associated with endoscopic findings suggestive of sinus ostial obstruction in subjects with allergic rhinitis. Measurement of nasal NO during humming may be a simple method to detect sinus abnormalities in these patients.

Goserelin versus danazol in the treatment of endometriosis.

Fourteen patients affected by endometriosis were treated by Danazol and by Goserelin. The two treatments were compared. From the study it emerged that both treatments were equally effective; however, the analogs gave a more favourable therapeutic profile.

Prognostic significance of the cyto-histological classification of infiltrating cervical epidermoid carcinoma.

One hundred-fivty-five cervical carcinomas were classified according to the Wentz and Reagan (21) cyto-histological system, and correlated to 5 year survival rates. This classification subdivides epidermoid carcinomas into three sub-groups: keratinizing carcinoma, large cell carcinoma, and small cell carcinoma. No significant differences between one group and another were observed. The importance of the type of therapy in conditioning the significance of this classification system is discussed.

Malignancy grading in squamous carcinoma of uterine cervix treated by surgery.

Some morphological patterns (histological type, vascular invasion, depth of invasion, lymphocytic infiltrate, mode of spread, necrosis) in 125 cases of squamous cervical carcinoma treated by surgery were analysed and graded in order to identify a histoprognostic score. Clinical data on F.I.G.O. stage, modality of surgical treatment, age, hormonal state (pre- or post-menopause) and 5-year survival were known for each patient. Two groups (low and high malignancy) were disclosed, and the difference of survival rate between the 2 was highly significant (P less than 0.001).

[Immunoglobulins and antibody titre in amniotic fluid of Rh-isoimmunized pregnant women (author's transl)]. / Immunoglobuline e titolo anticorpale nel liquido amniotico di gravide Rh-isoimmunizzate

On 90 samples of amniotic fluid coming from 54 cases of Rh-isoimmunization submitted to amniocentesis one or more times, the following tests were done: 1.) Coombs indirect test; 2) immunoelectrophoresis; 3) quantitative determination of immunoglobulins. The results were related to the degree of immunization as determined by Coombs indirect test on the mothers' serum and spectrophotometric curve of the amniotic fluid. For reasons of comparison, quantitative determination of immunoglobulins were performed on samples of amniotic fluid from 39 pregnant non-immunized patients. It was found that there was a direct relationship between antibody titers in the mothers' serum and those in the amniotic fluid, with the latter values always being inferior. Moreover, it was found that the level of IgG in the amniotic fluid and the degree of maternal immunization were proportional, with higher titers in the more severe cases. In addition to the routine tests, the determination of anti-Rh antibodies and titers of IgG in amniotic fluid can be useful in further evaluating degree of immunization in Rh-incompatibility. Furthermore, small quantities of IgA were found in amniotic fluid: these IgA were probably of secretory type (SIgA) and of amniotic origin, and therefore independent of any active immunization. Finally, all determinations of IgM were negative.