Mammalian Target Of Rapamycin (mTOR) inhibitors represent a new class of immunosuppressant drugs ... more Mammalian Target Of Rapamycin (mTOR) inhibitors represent a new class of immunosuppressant drugs extensively used for the prevention and the treatment of graft rejection in organ transplant recipients. Their current use is due to referred low nephrotoxic effects, particularly important in kidney transplanted and/or patients with renal failure. The most representative drugs of such class are Sirolimus (Siro) and Everolimus (Rad). Both drugs show a narrow therapeutic window, therefore, monitoring of whole-blood drug levels is recommended in order to optimize the therapy. Among the available assays, Liquid Chromatography coupled with UltraViolet or Electrospray Tandem Mass Spectrometry methods (LC/UV or LC/ESI-MSMS) are the most accurate and specific ones. A reliable alternative is represented by immunoassays, which offer the opportunity to minimize sample pre-treatment, thus reducing the time between drawing blood sample and measuring the drug concentration, an important aspect in high-throughput analyses. Despite this, a limitation in the use of immunoassays for therapeutic drug monitoring is the lower specifity compared with the chromatographic methods when analysing structurally-related drugs. New insights to optimize mTOR inhibitors regimens seem to be offered by the evaluation of CYP P4503A activity by using the probe drug approach. To such purpose, there are a number of major probe drugs used for in vivo studies including: midazolam, cortisol, lidocaine, nefedipine, dextromethorphan, erythromycin, dapsone and alfentanil. The aim of the present paper is to report the most recent knowledge concerning this issue, supplying a critical and comprehensive review for who is involved both in the clinical and analytical areas.
Many occupational toxicology studies have been focusing on the development of new specific biolog... more Many occupational toxicology studies have been focusing on the development of new specific biological indicators, owing to various factors: i) the exposure level decrement to hazardous agents in workplaces, ii) the biomarkers lack for numerous xenobiotics and for the risk evaluation following the exposure to complex mixtures, iii) the low availability of biologically effective dose indicators. Recent analytical and bioinformatics innovations, applied to omic sciences, allow an overall assessment of the cellular response to external stimuli, leading not only to the individuation of suitable biomarkers, but also, within the dynamic cell state, to the identification of predictive biomarkers related to pathologies induced by hazardous agents.Actually, the rapid development of analytical technologies, together with bioinformatics data management, allows a real change in biological monitoring aims. In fact, till now, biological monitoring studies investigated single events, being focused ...
Giornale italiano di medicina del lavoro ed ergonomia
In recent years, scientific literature has been giving more and more importance to the study of t... more In recent years, scientific literature has been giving more and more importance to the study of the occupational/environmental exposure to risk agents related to the onset of Amyotrophic Lateral Sclerosis (ALS), a neurodegenerative disease characterized by progressive muscular paralysis reflecting degeneration of motor neurons in the primary motor cortex. Aim of this work is to verify the state of art about the eventual role of occupational/environmental exposure to risk agents. Selected articles, on the basis of keywords, year of publication and topics, are related to occupational and environmental exposure to xenobiotics, and, in particular, to the exposure to heavy metals that could lead to neuronal damage mechanisms involved in ALS onset. The review shows that although the scientific production has increased the interest in the evaluation of extra-genetic causes of ALS onset, there are still few studies concerning the careful study of the work activities of the individual patien...
Many antineoplastic drugs (AD) commonly used in therapies display cytotoxic activity and are clas... more Many antineoplastic drugs (AD) commonly used in therapies display cytotoxic activity and are classified as potentially carcinogen , mutagen or teratogen. Since antineoplastic drugs are not selective towards malignant cells, their use may derive in the development of primary tumours in healthy subjects occupationally exposed to such agents, an occupational risk against which workers must be defended. The Environmental Monitoring represents a valid instrument in risk reduction strategies, since it allows the definition of the pollution levels actually present in the whole working areas and at the same time it can point out eventual structural and/or organizational lacks to be improved for pollution reduction. The present work reports results of Environmental Monitoring campaigns performed between 2004 and 2008 in oncology wards of different Italian hospitals. Analytes were chosen considering both their intrinsic toxicity properties and the amount handled for therapies. In particular, ...
Journal of occupational and environmental hygiene, Jan 20, 2015
Aircraft are constructed by modules that are covered by a "primer" layer, which can oft... more Aircraft are constructed by modules that are covered by a "primer" layer, which can often contain hexavalent chromium [Cr(VI)], known carcinogen to humans. While the occupational exposure to Cr(VI) during aircraft painting is ascertained, the exposure assessment of assembly workers (assemblers) requires investigations. Three biological monitoring campaigns (BM-I,II,III) were performed in an aviation industry, on homogeneous groups of assemblers (N=43) and controls (N=23), by measuring chromium concentrations in end-shift urine collected at the end of the working week and the chromium concentration difference between end- and before-shift urines. BM-I was conducted on full-time workers, BM-II was performed on workers after a 3-4 day absence from work, BM-III on workers using ecoprimers with lower Cr(VI) content. Samples were analyzed by atomic absorption spectroscopy and mean values were compared by T-test. Even if Cr concentrations measured during BM-I were lower than Biol...
Giornale italiano di medicina del lavoro ed ergonomia
Agricultural working contexts are characterized by numerous and heterogeneous working activities,... more Agricultural working contexts are characterized by numerous and heterogeneous working activities, leading to a noteworthy variety of risks for workers' healthcare. Safety and health risks vary according to each specific working activity, following seasonal cycles and productive process's phases. In particular, here the attention has been focused on the main risks factors present in large farms for fruit crops production, greenhouses and silos. Preventive measures for risks management are indicated in relation to each working activity step. Together with injury risks, essentially due to work equipment and machinery, unfavourable microclimatic conditions and hazards related to the manual handling of loads are often present, as well as chemical and biological risk agents adopted during the sterilization, disinfection and fertilization of grounds, plants and storage places. The reported findings highlight the necessity of increasing the risk consciousness of agricultural operato...
Doxorubicin (DOXO) is an anthracycline antibiotic which is used in the treatment of human maligna... more Doxorubicin (DOXO) is an anthracycline antibiotic which is used in the treatment of human malignancies such as leukemia, lymphoma and a number of solid tumors, particularly breast cancer. Anthracyclines have been reported to contaminate chemotherapy workstation surfaces as well as other workplaces surfaces. The occupational exposure to these drugs could occur in hospitals, for nurses involved in anthracyclines preparation and administration, in chemical industries during the commercial formulate syntheses, and in analytical laboratories. Numerous studies investigated cutaneous effects related to DOXO administration, on the contrary few literature data are available about effects on the skin due to the direct contact with the drug. The present study investigated the DOXO permeability of three commercially available gloves' types used to protect skin in occupational contexts, as well as the effects of DOXO on human keratinocyte cell line (HaCaT). The results suggest that the DOXO permeability of gloves depends not only on glove material but also on DOXO solutions' pH, in fact nitrile gloves can be penetrated by acid solutions, while neither natural rubbers nor nitrile gloves are permeable to neutral solutions. Moreover, DOXO solutions, even at low concentration, cause apoptosis in epithelial cells, through activation of intrinsic pathway p53-independent.
Ion trap mass spectrometry has been shown to be particularly suitable for the structural analysis... more Ion trap mass spectrometry has been shown to be particularly suitable for the structural analysis of high molecular weight peptides directly fragmented in the mass analyser without needing further sub-digestion reactions. Here we report the advantages of using multi-stage ion trap mass spectrometry in the structural characterisation of haemoglobin alkylated with epichlorohydrin and diepoxybutane. Alkylated globins were digested with trypsin and the peptide mixtures were analysed by MS(3). This technique allows the sequential fragmentation of peptides under analysis, giving rise to MS(3) product ion spectra with additional information with respect to MS(2) mass spectra. The results obtained complete the previously reported structural characterisation of alkylated haemoglobin, demonstrating the potential of ion trap mass spectrometry.
A specific and sensitive method based on gas chromatography/tandem mass spectrometry with on-colu... more A specific and sensitive method based on gas chromatography/tandem mass spectrometry with on-column injection was developed to quantify simultaneously cyclophosphamide and ifosphamide in urine by using trophosphamide as an internal standard. The urine samples were extracted with diethyl ether and derivatization was performed with heptafluorobutyric anhydride. The detection limits of cyclophosphamide and ifosphamide in urine samples were 0.1 and 0.5 ng ml(-1), respectively, with a signal-to-noise ratio of 3 : 1. The sensitivity, the specificity and the low cost of the instrumentation involved make this method suitable for economical analysis on a large scale, such as for the biological monitoring of occupational exposure to cyclophosphamide and ifosphamide in production plants and in hospitals during their pharmacological use.
Urinary benzene is used as biomarker of exposure to evaluate the uptake of this solvent both in n... more Urinary benzene is used as biomarker of exposure to evaluate the uptake of this solvent both in non-occupationally exposed population and in benzene-exposed workers. The quantitative determination of benzene in urine is carried out in a three steps procedure: urine collection, sample analysis by head space/solid phase microextraction/gas chromatography/mass spectrometry and analyte quantification. The adopted quantification method influences the initial step, hence the whole procedure. Two quantification approaches were compared as regards precision and accuracy: the calibration curves and the standard addition method. Even if calibration curves obtained by using urine samples from different subjects were always linear, their slopes and intercepts showed noteworthy variations, attributable to the influence of the biological matrix on benzene recovery. The standard addition method showed to be more suitable for compensating matrix effects, and a three-point standard addition protocol was used to quantify benzene in urine samples of 11 benzene-exposed workers (smokers and non-smokers). Urine from occupationally exposed workers was collected before and after work-shift. Besides urinary benzene, the applicability of the method was verified by measuring the urinary concentration of the S-phenylmercapturic acid, a specific benzene metabolite, generally adopted as biomarker in biological monitoring procedures. A similar trend of concentration levels of both analytes measured in urine samples collected before work-shift with respect to the after work-shift ones was found, showing the actual applicability of the standard addition method for biological monitoring purposes.
Mammalian Target Of Rapamycin (mTOR) inhibitors represent a new class of immunosuppressant drugs ... more Mammalian Target Of Rapamycin (mTOR) inhibitors represent a new class of immunosuppressant drugs extensively used for the prevention and the treatment of graft rejection in organ transplant recipients. Their current use is due to referred low nephrotoxic effects, particularly important in kidney transplanted and/or patients with renal failure. The most representative drugs of such class are Sirolimus (Siro) and Everolimus (Rad). Both drugs show a narrow therapeutic window, therefore, monitoring of whole-blood drug levels is recommended in order to optimize the therapy. Among the available assays, Liquid Chromatography coupled with UltraViolet or Electrospray Tandem Mass Spectrometry methods (LC/UV or LC/ESI-MSMS) are the most accurate and specific ones. A reliable alternative is represented by immunoassays, which offer the opportunity to minimize sample pre-treatment, thus reducing the time between drawing blood sample and measuring the drug concentration, an important aspect in high-throughput analyses. Despite this, a limitation in the use of immunoassays for therapeutic drug monitoring is the lower specifity compared with the chromatographic methods when analysing structurally-related drugs. New insights to optimize mTOR inhibitors regimens seem to be offered by the evaluation of CYP P4503A activity by using the probe drug approach. To such purpose, there are a number of major probe drugs used for in vivo studies including: midazolam, cortisol, lidocaine, nefedipine, dextromethorphan, erythromycin, dapsone and alfentanil. The aim of the present paper is to report the most recent knowledge concerning this issue, supplying a critical and comprehensive review for who is involved both in the clinical and analytical areas.
Many occupational toxicology studies have been focusing on the development of new specific biolog... more Many occupational toxicology studies have been focusing on the development of new specific biological indicators, owing to various factors: i) the exposure level decrement to hazardous agents in workplaces, ii) the biomarkers lack for numerous xenobiotics and for the risk evaluation following the exposure to complex mixtures, iii) the low availability of biologically effective dose indicators. Recent analytical and bioinformatics innovations, applied to omic sciences, allow an overall assessment of the cellular response to external stimuli, leading not only to the individuation of suitable biomarkers, but also, within the dynamic cell state, to the identification of predictive biomarkers related to pathologies induced by hazardous agents.Actually, the rapid development of analytical technologies, together with bioinformatics data management, allows a real change in biological monitoring aims. In fact, till now, biological monitoring studies investigated single events, being focused ...
Giornale italiano di medicina del lavoro ed ergonomia
In recent years, scientific literature has been giving more and more importance to the study of t... more In recent years, scientific literature has been giving more and more importance to the study of the occupational/environmental exposure to risk agents related to the onset of Amyotrophic Lateral Sclerosis (ALS), a neurodegenerative disease characterized by progressive muscular paralysis reflecting degeneration of motor neurons in the primary motor cortex. Aim of this work is to verify the state of art about the eventual role of occupational/environmental exposure to risk agents. Selected articles, on the basis of keywords, year of publication and topics, are related to occupational and environmental exposure to xenobiotics, and, in particular, to the exposure to heavy metals that could lead to neuronal damage mechanisms involved in ALS onset. The review shows that although the scientific production has increased the interest in the evaluation of extra-genetic causes of ALS onset, there are still few studies concerning the careful study of the work activities of the individual patien...
Many antineoplastic drugs (AD) commonly used in therapies display cytotoxic activity and are clas... more Many antineoplastic drugs (AD) commonly used in therapies display cytotoxic activity and are classified as potentially carcinogen , mutagen or teratogen. Since antineoplastic drugs are not selective towards malignant cells, their use may derive in the development of primary tumours in healthy subjects occupationally exposed to such agents, an occupational risk against which workers must be defended. The Environmental Monitoring represents a valid instrument in risk reduction strategies, since it allows the definition of the pollution levels actually present in the whole working areas and at the same time it can point out eventual structural and/or organizational lacks to be improved for pollution reduction. The present work reports results of Environmental Monitoring campaigns performed between 2004 and 2008 in oncology wards of different Italian hospitals. Analytes were chosen considering both their intrinsic toxicity properties and the amount handled for therapies. In particular, ...
Journal of occupational and environmental hygiene, Jan 20, 2015
Aircraft are constructed by modules that are covered by a "primer" layer, which can oft... more Aircraft are constructed by modules that are covered by a "primer" layer, which can often contain hexavalent chromium [Cr(VI)], known carcinogen to humans. While the occupational exposure to Cr(VI) during aircraft painting is ascertained, the exposure assessment of assembly workers (assemblers) requires investigations. Three biological monitoring campaigns (BM-I,II,III) were performed in an aviation industry, on homogeneous groups of assemblers (N=43) and controls (N=23), by measuring chromium concentrations in end-shift urine collected at the end of the working week and the chromium concentration difference between end- and before-shift urines. BM-I was conducted on full-time workers, BM-II was performed on workers after a 3-4 day absence from work, BM-III on workers using ecoprimers with lower Cr(VI) content. Samples were analyzed by atomic absorption spectroscopy and mean values were compared by T-test. Even if Cr concentrations measured during BM-I were lower than Biol...
Giornale italiano di medicina del lavoro ed ergonomia
Agricultural working contexts are characterized by numerous and heterogeneous working activities,... more Agricultural working contexts are characterized by numerous and heterogeneous working activities, leading to a noteworthy variety of risks for workers' healthcare. Safety and health risks vary according to each specific working activity, following seasonal cycles and productive process's phases. In particular, here the attention has been focused on the main risks factors present in large farms for fruit crops production, greenhouses and silos. Preventive measures for risks management are indicated in relation to each working activity step. Together with injury risks, essentially due to work equipment and machinery, unfavourable microclimatic conditions and hazards related to the manual handling of loads are often present, as well as chemical and biological risk agents adopted during the sterilization, disinfection and fertilization of grounds, plants and storage places. The reported findings highlight the necessity of increasing the risk consciousness of agricultural operato...
Doxorubicin (DOXO) is an anthracycline antibiotic which is used in the treatment of human maligna... more Doxorubicin (DOXO) is an anthracycline antibiotic which is used in the treatment of human malignancies such as leukemia, lymphoma and a number of solid tumors, particularly breast cancer. Anthracyclines have been reported to contaminate chemotherapy workstation surfaces as well as other workplaces surfaces. The occupational exposure to these drugs could occur in hospitals, for nurses involved in anthracyclines preparation and administration, in chemical industries during the commercial formulate syntheses, and in analytical laboratories. Numerous studies investigated cutaneous effects related to DOXO administration, on the contrary few literature data are available about effects on the skin due to the direct contact with the drug. The present study investigated the DOXO permeability of three commercially available gloves' types used to protect skin in occupational contexts, as well as the effects of DOXO on human keratinocyte cell line (HaCaT). The results suggest that the DOXO permeability of gloves depends not only on glove material but also on DOXO solutions' pH, in fact nitrile gloves can be penetrated by acid solutions, while neither natural rubbers nor nitrile gloves are permeable to neutral solutions. Moreover, DOXO solutions, even at low concentration, cause apoptosis in epithelial cells, through activation of intrinsic pathway p53-independent.
Ion trap mass spectrometry has been shown to be particularly suitable for the structural analysis... more Ion trap mass spectrometry has been shown to be particularly suitable for the structural analysis of high molecular weight peptides directly fragmented in the mass analyser without needing further sub-digestion reactions. Here we report the advantages of using multi-stage ion trap mass spectrometry in the structural characterisation of haemoglobin alkylated with epichlorohydrin and diepoxybutane. Alkylated globins were digested with trypsin and the peptide mixtures were analysed by MS(3). This technique allows the sequential fragmentation of peptides under analysis, giving rise to MS(3) product ion spectra with additional information with respect to MS(2) mass spectra. The results obtained complete the previously reported structural characterisation of alkylated haemoglobin, demonstrating the potential of ion trap mass spectrometry.
A specific and sensitive method based on gas chromatography/tandem mass spectrometry with on-colu... more A specific and sensitive method based on gas chromatography/tandem mass spectrometry with on-column injection was developed to quantify simultaneously cyclophosphamide and ifosphamide in urine by using trophosphamide as an internal standard. The urine samples were extracted with diethyl ether and derivatization was performed with heptafluorobutyric anhydride. The detection limits of cyclophosphamide and ifosphamide in urine samples were 0.1 and 0.5 ng ml(-1), respectively, with a signal-to-noise ratio of 3 : 1. The sensitivity, the specificity and the low cost of the instrumentation involved make this method suitable for economical analysis on a large scale, such as for the biological monitoring of occupational exposure to cyclophosphamide and ifosphamide in production plants and in hospitals during their pharmacological use.
Urinary benzene is used as biomarker of exposure to evaluate the uptake of this solvent both in n... more Urinary benzene is used as biomarker of exposure to evaluate the uptake of this solvent both in non-occupationally exposed population and in benzene-exposed workers. The quantitative determination of benzene in urine is carried out in a three steps procedure: urine collection, sample analysis by head space/solid phase microextraction/gas chromatography/mass spectrometry and analyte quantification. The adopted quantification method influences the initial step, hence the whole procedure. Two quantification approaches were compared as regards precision and accuracy: the calibration curves and the standard addition method. Even if calibration curves obtained by using urine samples from different subjects were always linear, their slopes and intercepts showed noteworthy variations, attributable to the influence of the biological matrix on benzene recovery. The standard addition method showed to be more suitable for compensating matrix effects, and a three-point standard addition protocol was used to quantify benzene in urine samples of 11 benzene-exposed workers (smokers and non-smokers). Urine from occupationally exposed workers was collected before and after work-shift. Besides urinary benzene, the applicability of the method was verified by measuring the urinary concentration of the S-phenylmercapturic acid, a specific benzene metabolite, generally adopted as biomarker in biological monitoring procedures. A similar trend of concentration levels of both analytes measured in urine samples collected before work-shift with respect to the after work-shift ones was found, showing the actual applicability of the standard addition method for biological monitoring purposes.
Uploads
Papers by N. Miraglia