Feasibility and efficacy of a multidisciplinary palliative approach in patients with advanced interstitial lung disease. A pilot randomised controlled trial.

INTRODUCTION AND OBJECTIVES: Interstitial lung diseases (ILDs) encompass a heterogeneous group of parenchymal lung disorders which have a significant burden on quality of life and exercise. The primary purpose of this randomised pilot trial performed in advanced ILD was to determine the feasibility and efficacy of a multidisciplinary palliative care approach (including physiotherapist, psychologist, pulmonologists, and palliative care doctors) to relieve patients' symptoms of dyspnoea, depression measured with the Center for Epidemiological Studies-Depression (CES-D) scale and quality-of-life (QoL) at 6 and 12 months. MATHERIALS AND METHODS: Fifty patients with confirmed interstitial lung disease at computed tomography (CT) scan and advanced disease were enrolled at our clinic. Patients were randomised to usual care group vs intervention group; in the intervention group, patients were scheduled to meet a physiotherapist, a psychologist, a palliative care doctor, and a pulmonologist specialized in ILD care. Data on dyspnoea, cough, quality of life and depression were recorded; patients in the intervention group were also tested to assess lower body flexibility and strength. RESULTS: Both groups showed a worsening in dyspnoea during the time course of the trial, but the Borg scale was less in the intervention group at 6 and 12 months. A similar trend was observed also for the CES-D scale. No differences were observed for the other scales. CONCLUSIONS: A multi-disciplinary palliative care intervention in patients with advanced fibrosing interstitial lung disease is feasible and effective. TRIAL REGISTRATION: NCT02929966 on ClinGovTrial.

Flow cytometry and capillary electrophoresis analyses in ethanol-stressed Oenococcus oeni strains and changes assessment of membrane fatty acid composition.

AIMS: This study aimed to investigate the dynamics and physiological heterogeneity of Oenococcus oeni under different conditions, cell membrane fluidity and permeability variations, and assessment of changes in cell surface charging rates. METHODS AND RESULTS: Flow cytometry, membrane fatty acid analysis and capillary electrophoresis were performed to study ethanol-induced variations. Different physiological states were assessed, revealing cell subpopulations able to adapt and withstand to environmental stress, in order to recover their functionality. Moreover, total results demonstrated changes in cell surface and membrane fatty acid redistribution with a saturation degree and an unsaturated/saturated fatty acid ratio fairly steady in control and in different ethanol stresses. CONCLUSIONS: This study revealed a great variability among O. oeni strains and the importance to investigate the mechanisms by a multiparametric approach based on the structural and physiological bacterial adjustments in different stresses tolerance. SIGNIFICANCE AND IMPACT OF THE STUDY: Intermediate physiological state assessment in O. oeni with recovery possibility could be an important criterion for potential starter culture application. The flow cytometry application with changes in monitoring membrane fatty acid composition and in surface charging rates allowed the characterization of sorted subpopulations that may contribute to further understanding of diversity and heterogeneity in physiology of bacterial populations.

The impact of method of biopsy on the incidence of breast cancer sentinel lymph node metastases.

BACKGROUND: The dislocation of the malignant cells along the needle tract during breast cancer (BC) diagnosis has been demonstrated by several studies. However, the published experiences that relate the diagnostic technique with sentinel node (SN) involvement are few and controversial. The aim of our analysis was to evaluate the impact of different techniques for preoperative BC biopsy among prognostic factors of metastases occurrence in SN. MATERIALS AND METHODS: We reviewed the institutional clinical database of our Center. A total of 674 patients were diagnosed between February 1999 and December 2006 with invasive BC. SN metastases classification followed the 2002 American Joint Committee on Cancer (AJCC) TNM pathological staging: macrometastases, micrometastases, isolated tumor cells or negative. Only macrometastases and micrometastases were considered positive. Concerning fine-needle aspiration cytology, we used disposable needles of the size of 21-27 G. For percutaneous biopsy we used cutting needle type "tru-cut"; the Gauge needle ranged between 14 and 20. RESULTS: At univariate analysis of specific parameters using positive SN as outcome, percutaneous diagnostic technique did not affect significantly the SN positivity (p = 0.60). At multivariate models only central quadrant lesion (p = 0.005) and lymph vascular invasion (LVI) presence (p < 0.0001) maintained the statistical significance as risk factor for positive SN status. Polytomic logistics models showed that only LVI maintained the statistical significance both for prediction of micrometastases and macrometastases. CONCLUSIONS: Our analysis showed that different techniques used for BC diagnosis did not influence SN status.

Minimal intervention dentistry: part 6. Caries inhibition by resin infiltration.

Resin infiltration has made possible an innovative way of treating initial carious lesions that fits perfectly with the concept of minimal intervention dentistry. Infiltration of carious lesions represents a new approach to the treatment of non-cavitated lesions of proximal and smooth surfaces of deciduous and permanent teeth. The major advantage of this method is that it is a non-invasive treatment, preserving tooth structure and that it can be achieved in a single visit. While this therapy can rightly be categorised as minimum intervention dentistry, clinical experience is limited and further controlled clinical trials are required to assess its long-term results. The inhibition of caries progression by resin infiltration should now be considered an alternative to invasive restorations, but involves early detection of lesions and does not allow for appropriate monitoring of the caries risk.

Minimal intervention dentistry: part 4. Detection and diagnosis of initial caries lesions.

The detection of carious lesions is focused on the identification of early mineral changes to allow the demineralisation process to be managed by non-invasive interventions. The methods recommended for clinical diagnosis of initial carious lesions are discussed and illustrated. These include the early detection of lesions, evaluation of the extent of the lesion and its state of activity and the establishment of appropriate monitoring. The place of modern tools, including those based on fluorescence, is discussed. These can help inform patients. They are also potentially useful in regular control visits to monitor the progression or regression of early lesions. A rigorous and systematic approach to caries diagnosis is essential to establish a care plan for the disease and to identify preventive measures based on more precise diagnosis and to reduce reliance on restorative measures.

Electrosynthesized poly(pyrrole)/poly(2-naphthol) bilayer membrane as an effective anti-interference layer for simultaneous determination of acethylcholine and choline by a dual electrode amperometric biosensor.

Several neural diseases appear related to the neurotransmitter acethylcholine (ACh) and its metabolite choline (Ch) brain levels so that their simultaneous determination is essential. A cross-talk and interference free dual electrode amperometric biosensor for the simultaneous determination of both analytes has been developed. Acetylcholinesterase (AChE) and choline oxidase (ChO) were immobilized by glutaraldehyde co-crosslinking with bovine serum albumin. A very efficient rejection of electroactive interferents has been achieved by a novel electrosynthesized polymeric bilayer membrane composed by overoxidised poly(pyrrole) and poly(2-naphthol) films. Sensitivities towards several electroactive interferents ranged from ca. 0.04% (e.g. ascorbate) to ca. 0.3% (e.g. dopamine) of those relevant to ACh and Ch (11 and 15 microA/microM, respectively). Detection limits (at S/N=3) in flow injection analysis were ca. 100 nM for both ACh and Ch at the ChO-AChE electrode and ca. 40 nM for Ch at the ChO sensor. Biosensor performances appear more than adequate for brain tissue homogenates and cerebrospinal fluids analysis where average levels in the low micromolar range are typically found.

[The stereotaxic core breast biopsy using the Mammotome: an alternative to intraoperative examination]. / La core-biopsy mammaria stereotassica con Mammotome: un'alternativa all'esame intraoperatorio.

The aim of this work was to evaluate the accuracy of vacuum-assisted biopsy by comparing it with frozen biopsy. 141 stereotaxic biopsies were performed by Mammotome (Ethicon Endo-Surgery, Hamburg) from January 2000 to March 2001. Biopsies were performed for microcalcifications (n = 105, 74.5%), irregular opacities (n = 20, 14.2%), regular opacities (n = 6, 4.2%), stellate lesions (n = 10, 7.1%). Histological analysis showed 85 (60.3%) benign lesions, 46 (32.6%) malignant lesions including (21 cases of carcinoma in situ and 25 invasive carcinomas) and 10 (7.1%) atypical lesions. All malignant lesions were subjected to surgery. In three cases (1 in situ and 2 invasive), core biopsy was excisional and no residual lesion was observed. Two of the carcinomas in situ revealed invasive features on the surgical biopsy. One of the atypical lesions was underestimated and the final diagnosis was "well differentiated carcinoma in situ." Only three of benign lesions underwent surgery after Mammotome biopsy. Among the 55 frozen-section biopsies of mammographically detected breast lesions performed in the same period, were one false-positive and 3 false-negative cases, while in 4 cases the diagnosis was deferred after paraffin embedding. Our results confirmed Mammotome biopsy as an effective alternative and a more reliable method than frozen-section examination.

A disposable amperometric biosensor for rapid screening of anticholinesterase activity in soil extracts.

A disposable amperometric biosensor for the determination of anticholinesterase activity in soil extracts is described. The sensitive membrane was obtained by co-crosslinking acetylcholinesterase and choline oxidase with bovine serum albumin using glutaraldehyde. The anticholinesterase activity of the soil extract was measured using chronoamperometry at 650 mV vs. Ag/AgCl to monitor the biocatalytically produced H2O2 before and after the inhibition step. An inhibition percentage of 38 +/- 4% was recorded for soil extracts spiked with 10 ppb of ethyl parathion. The device has the potential to be used as a gross sensor for the assessment of anticholinesterase activity in soil extracts.

An acetylcholinesterase/choline oxidase-based amperometric biosensors as a liquid chromatography detector for acetylcholine and choline determination in brain tissue homogenates.

A liquid chromatography (LC) detector based on a fast response and sensitive bienzyme amperometric biosensor for acetylcholine (ACh) and choline (Ch) is described. The detector fabrication consisted of glutaraldehyde co-crosslinking of acetylcholinesterase and choline oxidase with bovine serum albumin on the Pt working electrode of a conventional thin-layer electrochemical flow cell. The influence of some experimental parameters (e.g., enzyme loading, thickness of the bienzyme layer, flow rate) on the detector characteristics has been studied in order to optimize the analyte response while minimizing band-broadening and distortion. A mobile phase consisting of a phosphate buffer (I, 0.1 M; pH, 6.5) containing 5 mM sodium hexane sulfonate and 10 mM tetramethylammonium phosphate was found to give very satisfactory resolution and peak shape in ion-pair, reversed-phase LC. Linear responses were observed over at least four decades and absolute detection limits (at a signal-to-noise ratio of 3) were 12 and 27 fmol injected for Ch and ACh, respectively. After one month of intensive use in the LC system, the detector retained about 70% of its initial sensitivity. The potential of the described approach is demonstrated by the simultaneous determination of Ch and ACh in rat brain tissue homogenates.

Electrosynthesized non-conducting polymers as permselective membranes in amperometric enzyme electrodes: a glucose biosensor based on a co-crosslinked glucose oxidase/overoxidized polypyrrole bilayer.

A glucose amperometric biosensor based on glucose oxidase immobilized on an overoxidized polypyrrole (PPyox) platinum modified electrode, by glutaraldehyde co-crosslinking with bovine serum albumine, is described. The advantages of covalent immobilization techniques (e.g. high loading and long-term stability of the enzyme) are coupled with the excellent interferent rejection of electrosynthesized non-conducting polymers. The sensor showed an apparent Michaelis-Menten constant of 16 +/- 0.8 mM, a maximum current density of 490 microA/cm2 and a shelf lifetime of at least 3 months. Ascorbate, urate, cysteine and acetaminophen at their maximum physiological concentrations produced a glucose bias in the low micromolar range. Flow-injection response was linear up to 20 mM glucose with typical sensitivity of 84.0 +/- 1.5 nA/mM. The sensor was tested for glucose determination of untreated serum samples from both normal and diabetic subjects; results of amperometric assay compared well with those obtained by a standard enzymatic-colorimetric method.

[Quantitative assessment of pulmonary emphysema with computerized tomography. Comparison of the visual score and high resolution computerized tomography, expiratory density mask with spiral computerized tomography and respiratory function tests]. / Valutazione quantitativa dell'enfisema polmonare mediante tomografia computerizzata. Confronto tra il punteggio visivo con alta risoluzione nell'inspirazione, maschera della densità automatica con Tomografia Computerizzata spirale nell'espirazione ed esami funzionali respiratori.

CT is the most accurate method to detect pulmonary emphysema in vivo. We compared prospectively two different methods for emphysema quantitation in 5 normal volunteers and 20 consecutive patients with chronic obstructive pulmonary disease (COPD). All subjects were submitted to function tests and HRCT; three scans were acquired at preselected levels during inspiration. The type and extent of pulmonary emphysema were defined, using the time-honored visual score system, by two independent observers under blind conditions. Disagreements were subsequently settled by consent. All subjects were also examined with expiratory spiral CT, using a density mask program, at two different cut-off levels (-850, -900 HU). Visual score and expiratory spiral density mask values (-850 HU) were significantly correlated (r = 0.86), but the visual extent of emphysema was always higher than shown by expiratory spiral CT. The emphysema extent assessed with both CT methods correlated with the function result of expiratory airflow obstruction and gas diffusion impairment (visual score versus forced expiratory volume in one second: r = -0.81, versus single breath carbon monoxide diffusion: r = -0.78. Spiral expiratory density mask -850 HU versus forced expiratory volume in one second: r = -0.85, versus single breath carbon monoxide diffusion: r = -0.77). When -900 HU was used as the cut-off value for the expiratory density mask, the correlation with single breath carbon monoxide diffusion worsened (r = -0.56). Visual score and expiratory density mask -850 HU gave similar results and permitted COPD patients to be clearly distinguished from normal controls (p < 0.01). Residual lung volume, measured with expiratory spiral CT correlated significantly with residual volume measured with the helium dilution technique (r = 0.66), but CT values were always higher than function results. We believe the true residual volume should lie somewhere in between the CT value and the function results with the helium dilution technique and conclude that the extent of pulmonary emphysema can be confidently assessed with CT methods. Finally, the simple visual score may be as reliable as such highly sophisticated new methods as the spiral expiratory density mask. Expiratory studies offer new insights into different normal and abnormal features of COPD and respiratory impairment.

Role of palmitic acid on the isolation and properties of halorhodopsin.

Purified halorhodopsin was isolated from Halobacterium halobium as previously described (Duschl, A. et al. (1988) J. Biol. Chem. 263, 17016-17022). Two purple bands were eluted from phenyl-Sepharose column, indicating the presence of differently retained halorhodopsin forms; the absorption spectra of the two halorhodopsin bands in the dark were not different. By gas chromatography/mass spectrometry we could identify palmitate (which is only a minor lipid component of archaeal cells) among lipids associated with purple fractions. Typically the palmitate content of the first eluted band was higher than that of the second, indicating a correlation between the palmitate content and the retention time; from one to two fatty acid molecules per halorhodopsin molecule were present depending on the fraction analysed. Very little or no palmitate was released from denatured halorhodopsin. By adding palmitate to buffers used in the phenyl-Sepharose chromatography, only one sharp purple band was collected, corresponding to the less retained halorhodopsin fraction. Pentadecanoic fatty acid could also affect the halorhodopsin chromatography. Chromatography of halorhodopsin in the presence of beta-mercaptoethanol showed only one band, corresponding to the more retrained halorhodopsin form. The two halorhodopsin fractions had different photoreactivity; the less retained halorhodopsin fraction (at higher palmitate content) showed an higher rate of decay of the absorbance at 570 nm upon illumination. By following the decay of the absorbance at 570 nm upon addition of alkali in the dark, we found that the two halorhodopsin fractions had different pKa values of deprotonation.

An on-line semi-automated solid-phase extraction procedure for high-performance liquid chromatographic determination of lonidamine in serum.

A semi-automated solid-phase extraction procedure on-line with gradient elution reversed-phase chromatography permits the determination of lonidamine and its metabolite in human serum. The average recovery from serum at the 2.5 micrograms ml-1 level was (92.8 +/- 3.4)%. The limit of quantitation for a 100 microliter sample size was 50 ng ml-1. The within-day (n = 5) and between-day (n = 5) relative standard deviations for lonidamine determination in serum samples spiked at the 2.5 micrograms ml-1 level were 2.7% and 4.5%, respectively.

Effect of antidenaturant drugs on lysozyme deposit formation on soft contact lenses by liquid chromatography--electrochemical detection.

The effect of some antidenaturant drugs on the formation of lysozyme deposits on soft contact lenses was investigated by in vitro experiments. Results were obtained which encourage the use of this strategy to reduce lens spoilage. At the same time emphasis is given to the possibility that some functionalities (i.e. lysine groups) present in drug molecules could promote protein adsorption. In order to evaluate at a quantitative level the effectiveness of the drugs used to minimize the formation of deposits on soft contact lenses, a selective method for the determination of adsorbed lysozyme was developed. It is based on the hydrolysis of the 'lens/adsorbed lysozyme' system, followed by high performance liquid chromatography coupled to electrochemical detection of the released tryptophan residues. The lysozyme calibration curve was linear up to 5 mg ml-1 and the detection limit was 5 ng injected.

Solid-phase extraction of fluoropyrimidine derivatives on a copper-modified strong cation exchanger: determination of doxifluridine, 5-fluorouracil and its main metabolites in serum by high-performance liquid chromatography with ultraviolet detection.

A copper-modified strong cation exchange stationary phase was used for the solid phase extraction of the antineoplastic drug doxifluridine (5'-deoxy-5-fluorouridine) and some of its main metabolites from human serum. Sorption of fluoropyrimidine derivatives due to complex formation with copper(II) was maximal at pH values around neutrality. Analytes were eluted by ligand exchange with ammonia. The average recoveries ranged from 70% to 108%. Reversed-phase chromatography with ultraviolet detection was used for the separation and quantitation of the analytes. The overall procedure has been applied to serum samples from patients receiving doxifluridine chemotherapy, and a pharmacokinetic profile has been derived.

[The pediatrician and anorexia nervosa. Biological aspects]. / Il pediatra e l'anoressia mentale. Aspetti biologici.

Describing a case of anorexia nervosa the Authors review the clinical manifestations and organic alterations of this disease. The Authors point out the signs and symptoms which will usually suggest a diagnosis of anorexia nervosa. The course of the disease is monitored by the metabolic and hormonal analyses and the accompanying psychological features are underlined. The Authors present their therapeutic approach and emphasize the role of psychiatric treatment.

Simultaneous determination of pseudouridine and creatinine in untreated urine by ion-pair liquid chromatography with diode-array ultraviolet detection.

A simple procedure for the simultaneous determination of pseudouridine and creatinine in urine using ion-pair high-performance liquid chromatography with ultraviolet detection is described. It consists of simply diluting the filtered urine with mobile phase (1:20) followed by direct chromatographic injection. A single analysis takes only 10 min. This method has been applied to the analysis of urine samples from normal donors and patients with different types of cancer. The mean values, means, of the peak-area ratio of pseudouridine to creatinine were 61.79.10(-3) and 81.92.10(-3) for male and female normal donors, respectively. Out of twenty-five urine samples of patients with cancer examined, nineteen (all the forteen males included) had values higher than means + 2 sigma.

Determination of methotrexate in untreated body fluids by micellar liquid chromatography.

A fast and sensitive method for determination of the antineoplastic drug methotrexate in untreated serum and urine is described. Reversed-phase liquid chromatography with sodium dodecyl sulfate (SDS) micellar mobile phase has been used with direct sample injection. Changes in mobile phase variables such as SDS concentration and/or pH profoundly affected drug retention. Recovery was quantitative and the detection limit (90 nM for a 20 microL sample size) was below the range normally monitored. Multichannel UV detection provided strong evidence regarding peak purity and peak identity.