Biological monitoring and personal exposure to traffic-related air pollutants of elementary school-age children living in a metropolitan area.

An ever-growing burden of scientific evidence links air pollution to different aspects of human health even at very low concentrations; the impact increases for those living in urban environments, especially the youngest and the elderly. This study investigated the exposure to air pollution of urban school children of Milan, Italy, by personal and biological monitoring, in the frame of the MAPS-MI project. A total of 128 primary school children (7-11 years) were involved in a two-season monitoring campaign during spring 2018 and winter 2019. Personal exposure to airborne VOCs and eBC, and biological monitoring of urinary benzene (BEN-U) and methyl-tert-butyl ether (MTBE-U) were performed. Time-activity patterns, environmental tobacco smoke (ETS), spatial, and meteorological information were evaluated as determinants in mixed effects regression analysis. Children personal exposure was mostly quantifiable with median (5th-95th percentile) levels 1.9 (0.8-7.5) µg/m3 for eBC, and 1.1 (<0.6-3.4) and 0.8 (0.3-1.8) µg/m3 for benzene and MTBE, respectively; with values 2-3-fold higher in winter than in spring. In urine, median (5th-95th) BEN-U and MTBE-U levels were 44.9 (25.7-98.6) and 11.5 (5.0-35.5) ng/L, respectively. Mixed effect regression models explained from 72 to 93 % of the total variability for air pollutants, and from 58 to 61 % for biomarkers. Major contributors of personal exposure were season, wind speed, mobility- or traffic-related variables; biomarkers were mostly predicted by airborne exposure and ETS. Our results suggest that traffic-mitigation actions, together with parents' educational interventions on ETS and commuting mode, should be undertaken to lower children exposure to air pollution.

Exposure and Management of the Health Risk for the Use of Formaldehyde and Xylene in a Large Pathology Laboratory.

BACKGROUND: Formaldehyde and xylene are two hazardous chemicals widely used in pathology laboratories all over the world. The aim of this work was to survey a large volume pathology lab, measuring exposure of workers and residents to formaldehyde and xylene, and verify the efficacy of the undertaken preventive actions and the accomplishment with occupational limit values. METHODS: Environmental, personal, and biological monitoring of exposure to formaldehyde and xylene in different lab rooms and in 29 lab attendants was repeated yearly from 2017 to 2020. Continuous monitoring of airborne formaldehyde was performed to evaluate the pattern of airborne concentrations while specific tasks were performed. Several risk management and mitigation measures, including setting a new grossing room, reducing the number of samples to be soaked in formaldehyde, and improving the lab practices and equipment, such as the use of chemical hoods, were undertaken after each monitoring campaign, based on the results obtained from the exposure monitoring. RESULTS: Significant exposures to formaldehyde in pathologists and residents, especially during the grossing of samples, were observed in the first 2 years, with exposure exceeding the occupational exposure limit value; the following surveys showed that the risk management and mitigation measures were effective in reducing airborne concentrations and personal exposure. Xylene, assessed with both environmental and biological monitoring, was always well below the occupational exposure limit value and biological limit values, respectively. CONCLUSION: Critical exposure to air formaldehyde in attendants of a pathology laboratory could be reduced with the re-organization of lab spaces, new and improved work procedures, and awareness and training initiatives.

Environmental and biological monitoring of personal exposure to air pollutants of adult people living in a metropolitan area.

BACKGROUND: Human exposure to air pollutants, and specifically to particulate matter (PM) and volatile organic compounds (VOCs), may pose a relevant risk on human health. AIM: To evaluate the personal exposure of adults living and working in Milan (Italy) by environmental and biological monitoring. METHODS: Personal exposure of 51 volunteer adults to PM2.5, PM2.5-10 and selected VOCs, including benzene, toluene, ethylbenzene, o-xylene, m + p-xylene, methyl tert-butyl ether, naphthalene, hexane, cyclohexane, heptane, and limonene was assessed along a 24-h period via personal cascade impactors and radial diffusive samplers. Urine spot samples were collected to investigate the corresponding urinary biomarkers. Time-activity patterns were filled in by participants to explore the performed activities. Multiple regression models were applied to investigate the association between personal exposure, biomarker levels, and tobacco smoke, traffic exposure, commuting mode, cooking activities, and personal characteristics. RESULTS: Median personal exposure to PM2.5, PM2.5-10, benzene, toluene, ethylbenzene o-xylene, m + p-xylene, methyl tert-butyl ether, naphthalene, hexane, cyclohexane, heptane, and limonene were 36.1, 7.8, 2.3, 7.8, 2.1, 1.8, 4.7, 0.8, 0.3, 1.4, 2.5, 1.6, and 59.9 µg/m3, respectively. Median levels of urinary benzene, toluene, ethylbenzene o-xylene, m + p-xylene, naphthalene, hexane, and heptane were 78.0, 88.1, 21.5, 15.2, 43.9, 21.0, 11.0, and 22.5 ng/L, respectively. For personal exposure, multiple regression models explained up to 67% (PM2.5) and 61% (benzene) of variability, with major contribution from commuting mode and environmental exposure. For biological monitoring, multiple regression analysis explained up to 74% of urinary benzene, with a major contribution given by creatinine, and secondary contributions by commuting mode, personal exposure to airborne benzene and smoking. CONCLUSIONS: Personal exposure to air pollutants was lower than that measured in the past in Milan. Personal exposure was mainly driven by traffic variables, while internal dose was mainly driven by personal characteristics and smoking habit.

Development of a method to profile 2- to 4-ring polycyclic aromatic hydrocarbons in saliva samples from smokers and non-smokers by headspace-solid-phase microextraction-gas chromatography-triple quadrupole tandem mass spectrometry.

This study reports the development of a method based on headspace (HS)-solid-phase microextraction (SPME)-gas chromatography (GC)-triple quadrupole tandem mass spectrometry (MS/MS) for the quantification of 2- to 4-ring polycyclic aromatic hydrocarbons (PAHs) in saliva samples. Eight unmetabolized compounds (naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene and pyrene) were quantified using six deuterated PAHs as surrogate internal standards. The absence of matrix effect allowed saliva samples to be quantified by external calibration method. The optimized method resulted easy, with minimal sample pre-treatment (homogenization of the sample), and it achieved the highest sensitivity up to date: limits of quantification (LOQ) were in the 0.8-26.4 ng L-1 range, with a significant improvement in comparison with the few existing methods. Intra- and inter-run precisions provided CV values <18.1%, and accuracies within 20% of the spiked concentration. The application of the method to the analysis of fresh saliva samples collected by spitting from smokers (n = 10) and non-smokers (n = 10) showed that PAHs were quantifiable in all samples and that smokers had higher levels of all compounds than non-smokers. These results show that the method is suitable for quantifying low-boiling PAHs in saliva samples from individuals exposed at different PAH levels.

Urinary biomonitoring of subjects with different smoking habits. Part II: an untargeted metabolomic approach and the comparison with the targeted measurement of mercapturic acids.

BACKGROUND: Although thousands of different chemicals have been identified in cigarette smoke, the characterization of their urinary metabolites still requires significant research. The aim of this work was to perform an untargeted metabolomic approach to a pilot cross-sectional study conducted on subjects with different smoking habits and to compare the results with those of the targeted measurement of mercapturic acids. METHODS: Urine samples from 67 adults, including 38 non-smokers, 7 electronic cigarette users, and 22 traditional tobacco smokers were collected. Samples were analysed by liquid chromatography/time-of flight mass spectrometry. Data were processed using the R-packages IPO and XCMS to perform feature detection, retention time correction and alignment. One-way ANOVA test was used to identify different features among groups. Quantitative determination of 17 mercapturic acids was available from a previous study. RESULTS: One hundred and seventeen features, out of 3613, were different among groups. They corresponded to 91 potential metabolites, 5 of which were identified vs authentic standards, 43 were putatively annotated and 13 were attributed to chemical classes. Among identified compounds there were the mercapturic acids of acrolein, 1,3-butadiene, and crotonaldehyde; among putatively annotated compounds there were the glucuronide conjugated of 3-hydroxycotinine and the sulfate conjugate of methoxyphenol; with the lowest degree of confidence several sulfate conjugates of small molecules were annotated. Considering mercapturic acids, the coherence between the targeted and untargeted approach was found for a limited number of chemicals, typically the most abundant. CONCLUSIONS: Differences in the urinary levels of several compounds were associated to the different smoking habits, suggesting that the proposed approach is useful for the investigation of the metabolite patterns related to the exposure to toxicants. However, limitations were highlighted, in particular regarding the identification of low concentration compounds.

Urinary biomonitoring of subjects with different smoking habits. Part I: Profiling mercapturic acids.

BACKGROUND: While tobacco smoke contains thousands of chemicals, some of which are carcinogenic to humans, the content of electronic cigarette smoke is less known. This work aimed to assess and compare the exposure associated with different smoking habits by profiling urinary mercapturic acids as biomarkers of toxic compounds. METHODS: In this pilot study, sixty-seven healthy adults with different smoking habits were investigated: 38 non-smokers (NS), 7 electronic cigarette users (ECU), and 22 traditional tobacco smokers (TTS). Seventeen urinary mercapturic acids, metabolites of 1,3-butadiene (DHBMA, MHBMA), 4-chloronitrobenze (NANPC), acrolein (3-HPMA), acrylamide (AAMA, GAMA), acrylonitrile (CEMA), benzene (SPMA), crotonaldehyde (CMEMA, HMPMA), ethylating agents (EMA), methylating agents (MMA), ethylene oxide (HEMA), N,N-dimethylformamide (AMCC), propylene oxide (2-HPMA), styrene (PHEMA), and toluene (SBMA), were quantified, along with urinary nicotine and cotinine. RESULTS: Median urinary cotinine was 0.4, 1530 and 1772 µg/L in NS, ECU and TTS, respectively. Most mercapturic acids were 2-165 fold-higher in TTS compared to NS, with CEMA, MHBMA, 3-HPMA and SPMA showing the most relevant increases. Furthermore, some mercapturic acids were higher in ECU than NS; CEMA and 3-HPMA, in particular, showed significant increases and were 1.8 and 4.9 fold-higher, respectively. CONCLUSIONS: This study confirms that tobacco smoking is a major source of carcinogenic chemicals such as benzene and 1,3-butadiene; electronic cigarette use is a minor source, mostly associated with exposure to chemicals with less carcinogenic potential such as acrylonitrile and acrolein.

Biological Monitoring of Occupational Exposure to Metals in Electric Steel Foundry Workers and Its Contribution to 8-Oxo-7,8-Dihydro-2'-Deoxyguanosine Levels.

In this study, the urinary concentrations of selected metals in workers from an electric steel foundry in Tunisia were assessed and compared with existing biological limit values and general population reference values. Moreover, the association between oxidative DNA damage, measured as urinary 8-oxo-7,8-dihydro-2'deoxyguanosine (8-oxodG) and co-exposure to metals and polycyclic aromatic hydrocarbons (PAHs) was evaluated. Urinary levels of 12 metals were determined by inductively coupled plasma-mass spectrometry (ICP-MS) in end-shift spot samples from 89 workers. The urinary levels of phenanthrene (U-PHE), as marker of exposure to PAHs, and 8-oxodG were also available. Median levels ranged from 0.4 µg/L (cobalt, Co, and thallium, Tl) to 895 µg/L (zinc, Zn). Only 1% of samples was above the biological limit values for Co, and up to 13.5% of samples were above limit values for Cd. From 3.4% (Co) to 72% (lead, Pb) of samples were above the reference values for the general population. Multiple linear regression models, showed that manganese (Mn), Zn, arsenic (As), barium (Ba), Tl, and Pb were significant predictors of 8-oxodG (0.012 ≤ p ≤ 0.048); U-PHE was also a significant predictor (0.003 ≤ p ≤ 0.059). The variance explained by models was low (0.11 ≤ R2 ≤ 0.17, p < 0.005), showing that metals and PAHs were minor contributors to 8-oxodG. Overall, the comparison with biological limit values showed that the study subjects were occupationally exposed to metals, with levels exceeding biological limit values only for Cd.

ETS Exposure and PAH Body Burden in Nonsmoking Italian Adults.

Active smoking is associated with increased body burden of polycyclic aromatic hydrocarbons (PAHs); the aim of this study was to assess whether environmental tobacco smoking (ETS) increases the internal dose of PAHs. In 344 nonsmoking Italian adults, out of 497 individuals selected as representative of the population of the town of Modena, ETS exposure was evaluated by a self-administered questionnaire and by the measurement of urinary cotinine (COT-U). PAH exposure was assessed by the measurement of urinary 1-hydroxypyrene (1-OHPYR) and of ten urinary PAHs. In all subjects, median (5th⁻95th percentile) COT-U was 0.47 (.

Human biomonitoring of polycyclic aromatic hydrocarbonsand metals in the general population residing near the municipal solid waste incinerator of Modena, Italy.

BACKGROUND AND OBJECTIVES: A cross-sectional biomonitoring study was carried out to investigate exposure to incinerator emission in relation to the body burden of selected biomarkers in the population living around the plant. METHODS: Approximately 500 people, aged 18-69 yrs, living within 4 km from the incinerator were randomly selected form the population register. Exposure was measured through fall-out maps of particulate matter (PM), used as tracer for incinerator emissions. Ten metabolized polycyclic aromatic hydrocarbons (PAHs), from naphthalene to chrysene, 1-hydroxypyrene and twelve metals (Cd, Cr, Cu, Hg, Ni, Pb, Ni, Zn, V, Tl, As, Sn) were measured in spot urine samples. Confounders, such as diet, smoking, traffic, occupation and personal characteristics were assessed by questionnaires and objective measurements, and included into multivariate linear regression models. RESULTS: Metal concentrations in urine were in line with or higher than Italian reference limits, besides Cr and V with more than twofold concentrations. Metal levels did not show clear association to exposure categories. Most abundant PAHs were naphthalene (median 26.2 ng/L) and phenanthrene (7.4 ng/L). All PAHs, but benz[a]anthracene and 1-hydroxypyrene, were found in more than 52% of samples, and included in regression models. Significant associations between urinary PAHs and exposure were found, strong for fluorene, and weaker for naphthalene, fluoranthene and pyrene. Results were confirmed by sensitivity analyses. Correlation with variables reported in literature were observed. CONCLUSIONS: The study indicates that the emissions were very low and highlights that specific urinary PAHs provided useful information about the internal dose arising from incinerator emission.

Urinary 8-Oxo-7,8-Dihydro-2'-Deoxyguanosine in Tunisian Electric Steel Foundry Workers Exposed to Polycyclic Aromatic Hydrocarbons.

In this study, urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), as biomarker of oxidative DNA damage, was evaluated in Tunisian electric steel foundry workers and was associated with polycyclic aromatic hydrocarbon (PAH) exposure. Ninety-three healthy male workers were enrolled in the study; 8-oxodG was assessed by liquid chromatography-triple quadrupole mass spectrometry. Exposure to PAHs was evaluated by measuring 16 urinary PAHs (U-PAHs) and 8 monohydroxylated metabolites (OHPAHs). The median 8-oxodG level for all subjects was 3.20 µg l-1 (1.85 µg g-1 creatinine). No correlation between 8-oxodG and 1-hydroxypyrene or any other OHPAH was found. Significant linear correlations between 8-oxodG and some U-PAHs were found, particularly urinary acenaphthylene (r = 0.249), phenanthrene (r = 0.327), anthracene (r = 0.357), fluoranthene (r = 0.248), and pyrene (r = 0.244). Multiple regression analyses confirmed that urinary phenanthrene, anthracene, and naphthalene (the latter with a non-linear relationship) were predictors of 8-oxodG; job title, but not smoking, was a determinant of 8-oxodG; the variance explained by these models was up to 20%. The oxidative DNA damage assessed by urinary 8-oxodG was moderate and in the range of values reported in other occupational fields or in the general population. The results of this study indicate that the investigated biomarkers of PAH exposure were only minor contributors to urinary 8-oxodG.

Biological Monitoring of Occupational Exposure to Polycyclic Aromatic Hydrocarbons at an Electric Steel Foundry in Tunisia.

Occupational exposures during iron and steel founding have been classified as carcinogenic to humans, and the exposure to polycyclic aromatic hydrocarbons (PAHs) in this industrial setting may contribute to cancer risk. The occupational exposure to PAHs was assessed in 93 male workers at an electric steel foundry in Tunisia by biomonitoring, with the aims of characterizing the excretion profile and investigating the influence of job title and personal characteristics on the biomarkers. Sixteen 2-6 ring unmetabolized PAHs (U-PAHs) and eight hydroxylated PAH metabolites (OHPAHs) were analyzed by gas chromatography-triple quadrupole tandem mass spectrometry and liquid chromatography triple quadrupole tandem mass spectrometry, respectively. Among U-PAHs, urinary naphthalene (U-NAP) was the most abundant compound (median level: 643ng l(-1)), followed by phenanthrene (U-PHE, 18.5ng l(-1)). Urinary benzo[a]pyrene (U-BaP) level was <0.30ng l(-1) Among OHPAHs, 2-hydroxynaphthalene (2-OHNAP) was the most abundant metabolite (2.27 µg l(-1)). Median 1-hydroxypyrene (1-OHPYR) was 0.52 µg l(-1) Significant correlations among urinary biomarkers were observed, with Pearson's r ranging from 0.177 to 0.626. 1-OHPYR was correlated to benzo[a]pyrene, but not to five- and six-rings PAHs. A multiple linear regression model showed that job title was a significant determinant for almost all U-PAHs. In particular, employees in the steel smelter workshop had higher levels of high-boiling U-PAHs and lower levels of low-boiling U-PAHs than those of workers with other job titles. Among OHPAHs, this model was significant only for naphthols and 1-hydroxyphenanthrene (1-OHPHE). Smoking status was a significant predictor for almost all biomarkers. Among all analytes, U-PHE and 1-OHPHE were the less affected by tobacco smoke, and they were significantly correlated with both low- and high-molecular-weight compounds, and their levels were related to job titles, so they could be proposed as suitable biomarkers of PAH exposure at steel foundries. Based on 1-OHPYR levels, our findings show that occupational exposure of these workers was similar to that reported in recent studies of electric steel foundry workers. The multianalytic approach is useful in revealing different exposure levels among job titles.

MEN1-related hyperparathyroidism: response to cinacalcet and its relationship with the calcium-sensing receptor gene variant Arg990Gly.

OBJECTIVE: Primary hyperparathyroidism (PHPT) is a challenging problem in type 1 multiple endocrine neoplasia (MEN1) due to the high postsurgery recurrence rate. The aim was to evaluate the efficacy of cinacalcet in MEN1 patients in comparison with patients with sporadic PHPT (sPHPT) and the effect of Arg990Gly calcium-sensing receptor (CASR) polymorphism on the response to treatment. DESIGN: This is a randomized, crossover, double-blind study carried out in the University Hospitals. METHODS: Fifteen MEN1 patients with PHPT were randomized to two groups, one administered with 30 mg daily cinacalcet, titrated until calcium normalization, and one with placebo. After 3 months, patients were reassessed and after washout switched to the other treatment. For comparison, 20 sPHPT patients with similar calcium levels were administered with cinacalcet for 3 months. Ionized and total calcium, phosphate, and parathyroid hormone (PTH) were evaluated. CASR Arg990Gly was genotyped on blood DNA by direct sequencing. RESULTS: Cinacalcet normalized calcium, increased phosphate, and reduced PTH levels in all patients. Cinacalcet dosage required to normalize calcium in MEN1 and sPHPT was not significantly different (45±21 vs 54±25 mg/day). Few mild adverse events, not requiring drug withdrawal, were observed in both the groups. No association between Arg990Gly CASR polymorphism and response to cinacalcet was found. CONCLUSIONS: This short-term prospective study demonstrated that the efficacy profile of cinacalcet in patients with MEN1-related PHPT and in those with sPHPT was similar and was not influenced by the 990 CASR variant. Although long-term safety and efficacy data are required, cinacalcet might be considered a treatment option in MEN1 patients who have contraindications to surgery or persistent PHPT after surgery.

Filamin-A is essential for dopamine d2 receptor expression and signaling in tumorous lactotrophs.

CONTEXT: Dopamine agonists (DA) are the first choice treatment of prolactinomas. However, a subset of patients is resistant to DA, due to undefined dopamine D2 receptor (D2R) alterations. Recently, D2R was found to associate with filamin-A (FLNA), a widely expressed cytoskeleton protein with scaffolding properties, in melanoma and neuronal cells. OBJECTIVE: The aim of the study was to investigate the role of FLNA in D2R expression and signaling in human tumorous lactotrophs and rat MMQ and GH3 cells. DESIGN: We analyzed FLNA expression in a series of prolactinomas by immunohistochemistry and Western blotting. We performed FLNA silencing or transfection experiments in cultured cells from DA-sensitive or -resistant prolactinomas and in MMQ and GH3 cells, followed by analysis of D2R expression and signaling. RESULTS: We demonstrated reduced FLNA and D2R expression in DA-resistant tumors. The crucial role of FLNA on D2R was demonstrated by experiments showing that: 1) FLNA silencing in DA-sensitive prolactinomas resulted in 60% reduction of D2R expression and abrogation of DA-induced inhibition of prolactin release and antiproliferative signals, these results being replicated in MMQ cells that endogenously express FLNA and D2R; and 2) FLNA overexpression in DA-resistant prolactinomas restored D2R expression and prolactin responsiveness to DA, whereas this manipulation was ineffective in GH3 cells that express FLNA but not D2R. No alteration in FLNA promoter methylation was detected, ruling out the occurrence of epigenetic FLNA silencing in DA-resistant prolactinomas. CONCLUSIONS: These data indicate that FLNA is crucial for D2R expression and signaling in lactotrophs, suggesting that the impaired response to DA may be related to the reduction of FLNA expression in DA-resistant prolactinomas.

Influence of the d3GH receptor polymorphism on the metabolic and biochemical phenotype of GH-deficient adults at baseline and during short- and long-term recombinant human GH replacement therapy.

OBJECTIVE: A common polymorphic variant of GH receptor (exon 3 deletion, d3GHR) has been linked with increased response to recombinant human GH (rhGH) in some patients with or without GH deficiency (GHD). The aim of the study was to investigate the impact of the GHR genotype on the phenotype of GHD adults and on the metabolic effect of rhGH therapy. DESIGN: Prospective study of GHD patients evaluated before and during short- (1 year, n=100) and long-term (5 years, n=50) rhGH therapy. METHODS: Effects of rhGH on IGF1 levels, body composition (body fat percentage, BF%), body mass index, lipid profile, and glucose homeostasis (fasting insulin and glucose, insulin sensitivity indexes) were evaluated according to the presence or the absence of the d3GHR variant. RESULTS: The different genotype did not influence basal phenotype of GHD. Short-term rhGH determined normalization of IGF1 levels, decrease in BF%, and worsening of insulin sensitivity, independently from the presence of the d3GHR allele. A significant increase in high-density lipoprotein cholesterol occurred in the d3GHR group. Normalization of IGF1 levels and decrease in BF% were maintained after 5 years. Insulin sensitivity restored to basal values, though in d3GHR patients fasting glucose remained significantly higher than at baseline. After both 1 and 5 years, percentage of subjects with impaired glucose tolerance, similar in the two groups at baseline, decreased in fl/fl while doubled in d3GHR patients. In this last group, a long-term significant reduction in total and low-density lipoprotein cholesterol was also observed. CONCLUSION: The functional difference of d3GHR may influence some metabolic effects of rhGH on GHD adults.

Role of glucocorticoid receptor polymorphism in adrenal incidentalomas.

BACKGROUND: Adrenal incidentalomas (AI) have been associated with and an increased prevalence of metabolic and bone complications. The N363S and BclI polymorphisms of the glucocorticoid receptor (GR) have been associated with an increased sensitivity to glucocorticoid (GC). This observational study aims to evaluate whether BclI and N363S polymorphisms play a role in the development of complications in AI. MATERIALS AND METHODS: We enrolled 100 patients with AI (66 F; 34M). The presence of diabetes, arterial hypertension (AH), dyslipidaemia, osteoporosis and vertebral fracture (Fx), waist circumference and the Body Mass Index (BMI) were assessed. DNA samples were genotyped. Patients with wild-type BclI, wild-type N363S and heterozygous BclI polymorphism were classified as carriers of haplotype 1 (H1; n = 86), patients with homozygous BclI and heterozygous N363S polymorphism of GR of haplotype 2 (H2; n = 14). RESULTS: We found no clinical or biochemical differences between haplotype 1 and 2 groups, but a higher prevalence of the simultaneous presence of Fx plus AH in H2 patients (H2 n = 7, H1 n = 16, P = 0.01). Logistic regression analysis showed that the presence of Fx and of AH and the combination of the presence of Fx plus AH were associated with the H2 genotype regardless of the degree of cortisol secretion, age, BMI and BMD (OR 4.88, 95%CI 1.47-18.40, P = 0.05; OR 8.25, 95%CI 0.98-69.52, P = 0.05; OR 7.25, 95%CI 1.57-35.78, P = 0.011; respectively). CONCLUSIONS: In AI patients, the presence of the haplotype 2 of BclI and N363S is associated with the presence of AH, Fx and with the combination of Fx and AH.

d3-Growth hormone receptor polymorphism in acromegaly: effects on metabolic phenotype.

OBJECTIVE: A common polymorphic variant of the growth hormone receptor (GHR) is because of genomic deletion of exon 3 and has been linked with increased responsiveness to exogenous GH. The impact of this polymorphism in acromegaly, a disease characterized by endogenous excess of GH and partial loss of IGF-I feedback on tumoural GH secretion, is not clear. The aim of this study was to investigate possible influences of d3GHR on the GH/IGF-I relationship and metabolic parameters in acromegaly. DESIGN AND METHODS: Retrospective study on 76 acromegalic patients. Genotype analysis was carried out on leucocyte DNA by multiplex PCR assay. Clinical, hormonal and biochemical parameters at diagnosis were collected from patients' medical records. RESULTS: Forty-two patients (55.3%) were homozygotes for the allele encoding the full-length GHR (fl/flGHR), 27 patients were heterozygotes (fl/d3) and seven homozygotes (d3/d3) for the genomic deletion of exon 3. Heterozygotes and homozygotes for the d3 allele were considered together (d3GHR) and compared with fl/flGHR patients. d3GHR and fl/flGHR patients showed no difference in GH and IGF-I levels or in the relationship between these two parameters. Patients bearing d3GHR had a lower body mass index (BMI) than patients bearing fl/flGHR (25.8 +/- 2.1 vs. 28.1 +/- 4.8 kg/m(2), P < 0.05). Diabetes mellitus and hypertension were equally distributed, but more d3GHR patients had a normal glucose tolerance (66.7%vs. 56.3%, P < 0.05). The presence of d3GHR allele, and not BMI or age, was a significant negative predictor of insulin levels 120 min after oral glucose load (beta = -80.8, P < 0.05). CONCLUSIONS: This study supports the hypothesis that the d3GHR is functionally different from the fl/fl variant mostly for the effects on body weight regulation and on glucose metabolism.

The dopamine-somatostatin chimeric compound BIM-23A760 exerts antiproliferative and cytotoxic effects in human non-functioning pituitary tumors by activating ERK1/2 and p38 pathways.

The study investigated the effects of the dopamine-somatostatin chimeric compound BIM-23A760 on cell proliferation and apoptosis in cultured cells from human non-functioning pituitary tumors (NFPTs). Both BIM-23A760 and the dopaminergic agonist BIM-53097 induced a significant inhibition of cell proliferation associated with increased p27 expression, together with a significant increase in caspase-3 activity. Conversely, null or marginal effects were elicited by somatostatin analogs. Moreover, BIM-23A760 and BIM-53097 induced ERK1/2 and p38 phosphorylation and the blockade of these pathways prevented both the antiproliferative and the pro-apoptotic effects of these drugs. In conclusions the chimeric compound BIM-23A760 is able to exert cytostatic and cytotoxic effects in NFPTs, these phenomena being mainly mediated by DR2D and involving ERK1/2 and p38 pathways activation.

Effect of 9-cis retinoic acid on dopamine D2 receptor expression in pituitary adenoma cells.

The dopamine receptor subtype 2 (D2R) promoter contains a functional retinoic acid response element involved in the control of D2R expression. The aim of the study was to evaluate the effect of 9-cis retinoic acid (9-cis RA) on D2R protein expression in human pituitary adenomas and GH3 cell line. Treatment with 9-cis RA (100 nM for 48 hrs) caused a 109 +/- 32% increase of basal D2R levels in five of eight growth hormone (GH)-secreting adenomas (GH-omas), a 129 +/- 28% increase in 7 of 11 nonfunctioning adenomas, and no effect in two resistant prolactinomas by Western blotting. The lack of D2R induction in some tumors was not associated with a different pattern of retinoid x receptor (RXR) and retinoic acid receptor (RAR) isoform expression that was similar in all tumors by immunohistochemistry. While the induction of D2R did not affect the slight but significant inhibitory effect exerted by dopamine (10 nM) on in vitro GH release by GH-oma cultured cells, in pituitary GH3 cell lines cis-9 RA enhanced the dopamine-induced inhibition of in vitro GH release (% inhibition: 16 +/- 2 versus 26 +/- 5, P < 0.05), cell proliferation (25 +/- 2% versus 44 +/- 5%, P < 0.05) and cell viability (16 +/- 0.8% versus 29 +/- 1%, P < 0.05), likely by activating caspase-3 (28 +/- 3% versus basal, P < 0.05). In conclusion, this study provides novel evidence for a permissive role of retinoids on the expression of D2R in a good proportion of pituitary tumors and on the generation of pro-apoptotic signals in GH3 cell line.

4-Aminobiphenyl-hemoglobin adducts and risk of smoking-related disease in never smokers and former smokers in the European Prospective Investigation into Cancer and Nutrition prospective study.

The aim of this study was to evaluate whether biomarkers of environmental tobacco smoke exposure [i.e., 4-aminobiphenyl-hemoglobin (4-ABP-Hb) adducts] were predictive of the risk of tobacco-related cancers and diseases. We did a case-control study nested within the European Prospective Investigation into Cancer and Nutrition, involving 190 controls and 149 cases (incident cancer of the lung, bladder, pharynx, larynx, oral cavity, leukemias, and chronic obstructive pulmonary disease or emphysema deaths). All individuals were never smokers or ex smokers for >10 years. 4-ABP-Hb adducts were analyzed in peripheral blood collected before the onset of the disease (median, 7 years). Overall, 4-ABP-Hb adducts were higher, although not statistically significantly so, in cases (as a whole) than controls. In the control population, high fruit and vegetable consumption significantly lowered the frequency of detectable adducts (Fisher's exact test, P = 0.025). Restricting the analysis to women, 4-ABP-Hb adducts were higher in cases than controls (Mann-Whitney P = 0.036) and the odds ratio (OR) for the presence/absence of adducts was 2.42 [95% confidence interval (95% CI), 1.18-4.98]. Moreover, the association of adducts with the individual cancer types was stronger in women than in the whole study population, although statistically significant only for leukemias (OR, 2.77; 95% CI, 1.06-7.20). The results provide some evidence that women may be more susceptible to environmental tobacco smoke, as suggested by their higher adduct levels. The most important finding of this prospective study is that, at least in women, 4-ABP-Hb adducts may help identify subjects at high risk of cancers related to environmental tobacco smoke exposure.