A fast and validated mass spectrometry method for the evaluation of human serum albumin structural modifications in the clinical field.

A relatively fast analytical method for the identification and quantification of the post-transcriptional changes (PTCs) occurring in circulating human serum albumin (HSA) was developed. HSA is the most abundant protein in plasma and it represents the main determinant of plasma oncotic pressure, thus being the main modulator of fluid distribution between body compartments. Cirrhotic patients have low levels of HSA. Moreover, recent studies have demonstrated that during liver cirrhosis HSA presents PTCs affecting its properties. The HSA isoforms derived from these modifications could represent promising biomarkers for liver disease. Human plasma samples were collected from a cirrhotic patient (CH) and from an aged-matched non- cirrhotic subject (CT), purified by reverse-phase chromatography and analysed by an electrospray ionization quadrupole time-of-flight (ESI-Q-ToF) spectrometer. The deconvoluted ESI mass spectra from healthy subjects were all characterized by peaks attributed to mercaptoalbumin, nitrosylated, cysteinylated, glycated and N- terminal truncated HSA isoforms. The relative abundance of each isoform was derived and transformed into a relative per cent amount and the results were compared to those obtained analysing HSA from a CH plasma. The method was validated in terms of intra-day and inter-day reproducibility, both for quantitative results and PTCs molecular weight determination. The optimized method resulted in being effective in disclosing changes in HSA isoforms relative abundance and then it could be used for the systematic screening of cirrhotic patients to identify promising new biomarkers for liver diseases.

How new imaging techniques can aid in defining the cardiovascular profile of the high-risk patient.

Cardiovascular prevention has been developed in the last eight years producing an ever increasing amount of data requiring frequent updating. Studies using angiography to determine change in coronary obstruction have indicated progression, stabilization, or regression of coronary lesions associated with changes in plasma lipids and lipoproteins. Moreover, the guidelines on arterial hypertension published in 2007 listed the risk factors affecting prognosis but even by 2009 an update modified not only the list of risks, but even the philosophy behind the thought process which introduced as essential element in the prognosis of hypertension the ascertained existence of a damaged organ. Thus, the documentation of atherosclerotic vascular disease (plaques) and the quantification of its extension in the arterial tree became a determinant in the definition of cardiovascular risk. Magnetic Resonance (MRI) and coronary computed tomography (coro CT) applied to the heart and large vessels are the most promising methods.

[Bronchoscopy in respiratory intensive care]. / La broncoscopia in terapia intensiva respiratoria.

Born more than one century ago as a technique for the treatment of the respiratory emergencies due to foreign bodies inhalations, bronchoscopy plays nowadays a crucial and well-defined role in the management of the respiratory critical patient admitted in an intensive hospital setting thanks to its numerous applications both in the diagnostic and in the therapeutic fields. Despite the wide use of the bronchoscopy, especially of fibreoptic technique, it has never to be forgotten the importance of the operator's expertise specifically in this peculiar field of application in order to avoid usefulness risks and to prevent life-threatening consequences for the critical patient. The expert Pulmonologist represents the leader actor for the Clinical Governance of this branch of the Respiratory Intensive Care Medicine. In this review, the Authors examined the aspects connected with the choice of the instrument, the most clinical practice common indications, the risks and the consequences of the use of the bronchoscopy in respiratory intensive care.

[Pulmonary arterial hypertension due to COPD]. / Hypertension artérielle pulmonaire et bronchopathie chronique obstructive.

In this paper we review the current knowledge on pulmonary hypertension (PH) occurring in COPD. PH is defined as a mean pulmonary arterial pressure at rest greater than 20 mmHg measured by right heart catheterisation. PH is usually present during exercise before appearing at rest. PH in COPD increases the risk of hospitalisation and darkens the disease prognosis. Chronic hypoxemia is the major contributor to PH, but remodelling of arterial wall and mechanical factors such as hyperinflation also play a role. Transthoracic echocardiography is the most useful non invasive investigation, but right heart catheterisation is necessary to ascertain the diagnosis. Long term 02 supplementation is the basis of the treatment while vasodilatators may worsen hypoxemia.

Histone proteins determined in a human colon cancer by high-performance liquid chromatography and mass spectrometry.

The application of reversed-phase high-pressure liquid chromatography under gradient conditions and electrospray ion trap mass spectrometry (LC-ESI-MS) to the analysis of global modification levels of core histones is described. The optimised LC-ESI-MS method was applied for the first time to the characterisation of histones extracted from HT29, a human colon cancer cell line. Eight histones (H1-1, H1-2, H2A-1, H2A-2, H2B, H3-1, H3-2, H4) were separated on a C4 stationary phase with complete resolution, never reached in previous HPLC-MS methods, by using a gradient elution with the combined presence of heptafluorobutyric acid and formic acid as acidic modifiers in the mobile phase. Heptafluorobutyric acid was found to improve selectivity, whereas the presence of formic acid decreased ion suppression. Histones eluted from the column were detected with an ion trap mass spectrometer with an electrospray source. The peak averaged mass spectra were reconstructed by Mag Tran 1.0 software and the mass of the various isoforms of histones were derived. Method validation was conducted by performing the same sample analysis by coupling LC-ESI to a quadrupole-time-of-flight mass spectrometer (Q-TOF). The number of histone forms and their mass were found to differ not significantly from those obtained by ion trap mass spectrometer. Also the relative modifications abundance within the same histone type was found following the same trend as the two mass analysers. This method was then applied to the characterisation of changes in histone modification in HT29, never analysed by LC-MS before, treated with histone deacetylase inhibitors such as valproate and sodium butyrate, also used in preclinical trials as anticancer drugs. In particular, both the inhibitors produced a significant increase in H4 histone acetylated forms: 89% increase of the diacetyl dimethyl H4 form was observed with 1mM valproate supplementation, whereas 5 mM butyrate led to a 68% increase of the same form. Triacetyl monomethyl H4 (11,377 Da) and triacetyl dimethyl H4 (11,390 Da) were found only in cells treated with butyrate. Selective changes of H3 histone were detected with butyrate, in agreement with recently reported western blotting studies. Modifications in the H2A histone degree of acetylation were revealed by treatment of the cells with butyrate (H2A-1, H2A-2) and valproate (H2A-2). The results of the proposed methodology confirmed that inhibition of histone deacetylases caused histone hyperacetylation, responsible for decondensation and reorganization of interphase dynamic chromatin. This method resulted in selective and sensitive method to monitor variations in the acetylation and methylation state of histones after treatment of HT29 with inhibitors, and is therefore suitable for further application in new drug discovery for tumour therapy.

Non-invasive positive pressure ventilation in acute hypercapnic respiratory failure: clinical experience of a respiratory ward.

BACKGROUND: Although a controlled trial demonstrated that non-invasive positive pressure ventilation (NIV) can be successfully applied to a respiratory ward (RW) for selected cases of acute hypercapnic respiratory failure (AHRF), clinical practice data about NIV use in this setting are limited. The aim of this observational study is to assess the feasibility and efficacy of NIV applied to AHRF in a RW in everyday practice. METHODS: Twenty-two percent (216/984) of patients consecutively admitted for AHRF to our RW in Arezzo (years: 1996-2003) received NIV in addition to standard therapy, according to pre-defined routinely used criteria. Tolerance, effects upon arterial blood gases (ABG), success rate (avoidance a priori criteria for intubation) and predictors of failure of NIV were analysed. RESULTS: Nine patients (4.2%) were found to be intolerant to NIV, while the remaining 207 (M: 157, F: 50; mean (SD) age: 73.2 (8.9) yrs; COPD: 71.5%) were ventilated for >1 hour. ABG significantly improved after two hours of NIV (pH: 7.32 (0.06) versus median (Interquartiles) 7.28 (7.24-7.31), p<0.0001; PaCO2: 71.9 (13.5) mmHg versus 80.0 (15.2) mmHg, p<0.0001; PaO2/FiO2: 212 (66) versus 184 (150-221), p<0.0001). NIV succeeded in avoiding intubation in 169/207 patients (81.6%) with hospital mortality of 15.5%. NIV failure was independently predicted by Activity of Daily Living score, pneumonia as cause of AHRF and Acute Physiology and Chronic Health Evaluation III score. CONCLUSIONS: In clinical practice NIV is feasible, effective in improving ABG and useful in avoiding intubation in most AHRF episodes that do not respond to the standard therapy managed in an RW adequately trained in NIV.

Gardening work and heavy metals in urban environment.

Urban soil may be a source of occupational exposure to various pollutants in gardening and land cultivation. This paper presents data of a one-year follow-up of lead, cadmium, nickel, chromium, and vanadium in the environment of the city of Bologna. Samples of soil and leaves were collected at three locations; gardens from the inner-city high-traffic area, parks in moderate-traffic area, and parks in suburban, low-traffic area. The top and deeper layers of soil and leaves were mainly polluted by lead at all locations, which corresponded to the traffic density. Personal samplers recorded greater concentrations of airborne metals than did the area samplers but the values kept below the threshold limit established by the American Conference of Governmental Industrial Hygienists for the working environment. Due to cumulative nature and interactive effects of toxic metals with other toxic and essential elements, long-term exposure to metals in the urban environment may be a health risk for occupationally exposed gardeners.

Mortality among workers in an Italian cigarette factory.

A cohort study was conducted to evaluate the mortality pattern among female and male workers in a cigarette factory. The study followed 972 female workers and 761 male workers with at least 6 months of cumulative employment from 1 January 1962 through 1 July 1996. Among women, mortality from all causes of death [standardized mortality ratio (SMR) = 0.9; 95% confidence interval (CI) = 0.8-1.1] and mortality from all malignant neoplasms (SMR = 1.1; CI = 0.9-1.3) were consistent with reference rates. Male workers had a significantly reduced overall mortality (SMR = 0.8; CI = 0.7-0.9), while mortality from all malignant neoplasms was as expected (SMR = 0.9; CI = 0.7-1.0). Among female workers the frequencies of deaths from diseases of nervous system (SMR = 2.0; CI = 1.1-3.4) and from non-Hodgkin's lymphoma (NHL) (SMR = 2.7; CI = 1.0-5.6) were elevated at a statistically significant level. No association between duration of employment and diseases of nervous system was observed. A higher risk for NHL, based on three deaths, was reported among female workers with 15 or more years of employment (SMR = 8.1; CI = 2.2-21.0). Although based on small numbers, the excess of NHL here reported suggests that potential exposure to foliar residues of pesticides should be thoroughly considered in tobacco manufacturing.

Endobronchial granular cell tumour.

Granular cell tumour of the lung is a rare benign tumour. We report a case of endobronchial granular cell tumour which occurred in a 70-yr-old male with haemoptysis. Because of the size of the tumour (greater than 8 mm) and the associated pseudoepitheliomatous hyperplasia with marked atypia, surgical resection was performed. No neoplastic relapse has been observed in nearly 4 yrs of follow-up.

Use of ultrasonography in monitoring work-related carpal tunnel syndrome: a case report.

Carpal tunnel syndrome (CTS) is a syndrome whose diagnosis is well established. One cause could be occupational factors, while others have no relation to work or the work environment. We present in this article a case report regarding a worker affected by CTS, which is of interest concerning the sensitivity of ultrasonography and electroneurography, applied as diagnostic methods, related in our protocol to the variations in occupational exposures. The case reports an agricultural worker, whose tasks required repetitive and high frequency movements of the hand-arm. Diagnosis of CTS used ultrasonography and electroneurography techniques. In our opinion, the clinical evolution of CTS encompasses three "work-related" phases (preclinical phase; phase of nerve compression; phase of irreversible damage). Ultrasonography provides greater information about the evolution of CTS, as well as other cumulative trauma disorders, and is able to discern tendinitis of flexors causing a compression on the median nerve in the carpal tunnel.

[Non-invasive nasal ventilation in a a case of hypercapnic coma]. / La ventilazione non invasiva per via nasale in un caso di coma ipercapnico.

Noninvasive positive pressure ventilation (NPPV) via nasal mask is well known to be effective in the treatment of acute respiratory failure (ARF) secondary to chronic obstructive pulmonary disease (COPD). A case of ARF with hypercapnic coma due to exacerbation of COPD is described. Six hours of conservative therapy with oxygen and medical treatment did not show any result. As endotracheal intubation (ET) was avoided on the basis of advanced age, poor life expectancy of the patient and family wish, NPPV was set up using a pressure triggered ventilator. After 61 hours of uninterrupted NPPV, the acid-base alteration and the lethargic status was fully reversed. The conclusions is drawn that NPPV may be useful also in the treatment of patients affected by severe decompensated hypercapnic respiratory failure in whom ET is not indicated.

Cumulative trauma disorders of the upper limbs in workers on an agricultural farm.

The work associated with repetitive efforts and inadequate resting periods, strong physical exertion, awkward postures or static positioning exposes workers to the risk of cumulative trauma disorders of the upper limbs. These risk factors are present in many agricultural activities. A study was carried out among workers on an agricultural farm. The workers' histories were taken and they were given periodical medical check-ups. The presence of upper limb disorders was shown in a group of workers. A sample of 42 people was selected for the study by means of specific tests: electromyography, ultrasonography and laser-doppler flowmetry. The tests showed a high incidence of carpal tunnel syndrome and microcirculation disorders. The study confirmed that electromyography, ultrasonography and/or laser-doppler flowmetry are highly useful tools for identifying cumulative trauma disorders.

gamma-Glutamyltransferase isoenzyme pattern in workers exposed to tetrachloroethylene.

In compliance with the mandatory medical surveillance of workers exposed to tetrachloroethylene (PCE) in Italy, isoenzyme fractioning of serum gamma-glutamyltransferase (GGT) was performed on 141 workers of both sexes and on 130 control subjects. None of the workers showed any clinical symptoms of liver disease and their enzymatic profiles, including AST, ALT, 5'-NU, ALP, and GGT, were within the normal reference limits. A statistically significant increase in total GGT serum level was found in the exposed subjects, which was associated with an increase in one of the two fractions normally present in healthy individuals (GGT-2), as well as with the appearance and progressive increase of the level of a fraction (GGT-4) considered to be an expression of hepato-biliary impairment. Further research is ongoing among these workers, which will clarify whether or not electrophoretic GGT tests may be useful in detecting liver function changes due to occupational exposure to PCE.

Contribution of peripheral blood pooling to central hemodynamic disturbances during endotoxin insult in intact dogs.

The aim of the present study was to determine possible effects of Escherichia coli endotoxin on peripheral vascular compliance and relate them to concomitant central hemodynamic disturbances. Endotoxin was infused at 0.25 micrograms/kg.min during 2 h in six anesthetized dogs, while six additional animals served as controls. Vascular compliance of the systemic circulation was calculated in intact animals from the changes in CVP after known changes in systemic blood volume. In control dogs, vascular compliance averaged 2.3 ml/mm Hg.kg body weight. During slow endotoxin infusion, cardiovascular effects were measurable only after a certain period of time had elapsed from the start of endotoxin insult and consisted of hypotension associated with systemic vasodilation. Systemic BP decreased gradually from 124 to 68 mm Hg while vascular compliance was finally increased by 100%, when compared to control values. This latter rise was responsible for a reduction in the cardiac preloads. Pulmonary wedge pressure and CVP were decreased from 7.1 to 3.4 and from 4.5 to 2.6 mm Hg, respectively. However, parallel to the decrease in left ventricular preload, endotoxin induced a progressive decrease in left ventricular afterload. Because of the balance in ventricular loading, cardiac output remained almost unchanged. After volume loading (dextran 30 ml/kg), cardiac output was remarkably increased from 3.28 to 6.24 L/min.m2 while peripheral vasodilation was not affected by this maneuver. It is concluded that low dose endotoxin infusion induces in dogs a hemodynamic pattern similar to human sepsis. The left ventricular loading changes are related to an enhanced systemic vascular compliance from 2.3 to 4.5 ml/mm Hg.kg. High flow shock state is encountered provided peripheral blood pooling is compensated by adequate volume replacement.

[Electromyography of those exposed to vibration]. / Elettromiografia negli esposti a vibrazioni.

Electrophysiological findings in the median and ulnar nerves of a group of ten workers who operate hand-held vibrating power tools are described. Motor conduction velocities, motor latencies and amplitude of evoked compound muscle action potentials were measured. We have observed statistically significant changes in the distal latencies and in the amplitude of evoked compound muscle action potentials of median nerves in all the subjects examined, whereas the motor conduction velocities were normal in both median and ulnar nerve. These findings are compatible with functional changes without a clinically manifest neuropathy and they are suggestive of a compression damage such as carpal tunnel syndrome: the possible pathogenic mechanism of these damages are discussed. It's necessary to emphasize the importance of further studies in order to estimate the type and entity of the exposure to vibrations to show a cause-effect relationship.