[Influence of occupational exposures in nonmuscle infiltration bladder cancer: preliminary results of a prospective study from October 2005 to February 2009]. / Influence des expositions professionnelles dans les cancers n'infiltrant pas le muscle détrusor. Résultats préliminaires d'une étude prospective d'octobre2005 à février2009.

UNLABELLED: Bladder cancer is a common condition in industrialized countries. If tobacco is still the main risk factor in lung cancer, occupational exposures carcinogens should not be underestimated. GOAL: The significant excess of bladder cancer in the north part of France, with high manufacture concentration likely to have employees exposed to bladder carcinogens, has led us to study the influence of these exposures in the natural history of bladder cancer. PATIENTS AND METHODS: We prospectively conducted a descriptive case-control study. A questionnaire was developed by the department of occupational disease and clinical, radiological, histological, therapeutic data were registered at the University Hospital of Lille. From October 2005 to February 2009, 69 patients were included in the study, 37 exposed to occupational carcinogens and 32 in the control group. RESULTS: Mean age was 61.6 years vs. 61.8 years and the sex ratio of 7.33 men to one woman vs. one woman for three men respectively in the two groups. The average age of patients exposed to polycyclic aromatic hydrocarbons was 59.7 years. Smokers were 86.5% and 87.5% respectively. Follow-up was 38.4 and 32.9 months respectively. Nonmuscle invasive bladder cancer were more frequent (P=0.019) in the exposed group (84.4%) than in the unexposed group (67.8%) even after adjustment for smoking (P=0.0142). The histological type, grade, presence of CIS, the early recurrence at 3 months, the number of lesions at diagnosis does not differ in the two groups even after adjustment for smoking or after subgroup analysis of the most frequent exposure. The exposure to polycyclic aromatic hydrocarbons (62%) and aromatic amines (37.8%) were the most represented. Of 37 patients, 13 (35%) were making a statement as an occupational disease (eight according to Table 15 ter, two according to Table 16 bis and two presented to IRB). To date one single patient is recognized as an occupational disease. CONCLUSION: We did not observe any worsening of the prognosis of bladder cancer following occupational carcinogen exposure except for the mean age at diagnosis. The small size of the population studied and the importance of smoking partner have hampered the analysis of occupational exposures.

Cytomegalovirus seroprevalence in exposed and unexposed populations of hospital employees.

The objective of this work was to compare the seroprevalence of cytomegalovirus in an unexposed and exposed population, both working in a hospital, and to study the occupational risk factors related to seropositivity, while taking personal risk factors into account. We conducted a cross-sectional study in a French hospital over a period of 12 months. The overall seroprevalence among the 550 subjects was 49.5%. The multivariate analysis showed that seropositivity was significantly associated with age (36-43 years: odds ratio [OR] = 1.7; 95% confidence interval [CI]: [1.1-2.8]) and working as a pediatric nurse's aide (OR = 1.8; 95% CI: [1.1-2.8]). This study confirms the need to improve prevention procedures in the workplace, including screening, information, and hygiene rules.

Age-dependency of mycophenolate mofetil dosing in combination with tacrolimus after pediatric renal transplantation.

BACKGROUND: Dosing of mycophenolate mofetil (MMF) must be lower in combination therapy with Tacrolimus (Tac) than with Cyclosporine. One study with mostly adolescent recipients recommended an MMF dose of 250 mg/m2 BID. Because this dose resulted in low area-under-the-curve (AUC) in our infant population, we retrospectively analyzed all available pharmacokinetic (PK) profiles in pediatric renal transplant patients on MMF plus Tac therapy to propose appropriate MMF dosing in pediatric patients of all ages. PATIENTS AND METHODS: Forty-four PK profiles were performed in 27 patients (median age, 11.6 years; range, 1.8-20.7 years). The investigations were performed at a median of 299 days (range, 24-3424) after transplantation. Ten patients were converted to Tac plus MMF, all others received this as primary therapy. For patients with repeated measurements, we calculated the average AUC and doses. We used first-order PK modeling to calculate the doses for a mycophenolic acid (MPA) AUC of 60 ug*h/mL and a Tac AUC of 150 ng*h/mL. RESULTS: The mean Tac dose was 2.6 +/- 1.2 mg/m2/d or 0.086 +/- 0.038 mg/kg/d, resulting in an average AUC of 120.6 +/- 30.4 ng*h/mL. The MMF dose was not normally distributed; the median dose was 549 mg/m2/d (range, 146-1413) and the median MPA AUC was 49.8 ug*h/mL (range, 26.7-156.0). The mean dose for a Tac AUC of 150 ng*h/mL was 3.50 +/- 1.77 mg/m2/d (0.117 +/- 0.058 mg/kg) and was independent of age or time after transplantation. By contrast, we found a negative relationship between the dose per m2 (r2 = 0.29; P = 0.0038) or per kg (r2 = 0.58; P < .0001) required for an MPA AUC of 60 ug*h/mL and patient age. Converted and primary patients behaved identically. The dosing requirement decreased from 500 mg/m2 BID in 2-year-old patients to 250 mg/m2 in adolescents. There was substantial interpatient variability of 44%. CONCLUSIONS: Higher MMF doses are required for young children. Our data suggest a starting dose for infants of 500 mg/m2 BID, with PK monitoring of MPA due to substantial interpatient variability.

Universal approach to pharmacokinetic monitoring of immunosuppressive agents in children.

Current data indicate that pharmacokinetic (PK) monitoring of cyclosporin microemulsion (CsA) should be performed using the 2-h concentration (C2), that tacrolimus (Tac) is commonly monitored using the trough level, and mycophenolate mofetil (MMF) should be monitored using the 1-h (C1), 2-h (C2) and 6-h (C6) concentrations. The three differing time-point requirements are cumbersome, and we aimed to develop universal guidelines for all three drugs using a large number of full PK profiles in children. One-hundred and twenty two stable pediatric patients, receiving either CsA (165 PK profiles, 69 patients, 24 with concomitant MMF) or Tac (122 PK profiles, 53 patients, 18 with MMF) were analyzed retrospectively. Pearson r for the CsA C2 was 0.90 [95% confidence interval(CI): 0.86-0.92], for Tac C2 r was 0.86 (95% CI: 0.80-0.90), and for MPA C2 r was 0.77 (95% CI: 0.68-0.83), respectively. For MPA, at least three time-points are required to accurately estimate the area under the concentration-time curve (AUC), and C1, C2 and C6 serve as best markers. Excellent AUC estimations could be obtained from a limited sampling strategy from C1, C2 and C6 or C0, C1, C2 and C4 with clinically acceptable errors for all three drugs. The AUC can be estimated with great precision by using an identical approach for all three drugs. Target AUCs for a given time-point after transplantation remain to be established.

Effect of cyclosporine on mycophenolic acid area under the concentration-time curve in pediatric kidney transplant recipients.

Mycophenolic acid (MPA), the active compound of mycophenolate mofetil (MMF), shows substantial interindividual and intraindividual variability. It was recently shown that in vitro calcineurin inhibitors alter the bioavailability of MPA by dose-dependent inhibition of MPA glucuronidation. The authors retrospectively analyzed full 10-point profiles for both MPA and cyclosporine (CyA) in 23 pediatric patients receiving MMF and cyclosporine microemulsion (Neoral; Novartis Pharmaceuticals Canada; Dorval, Quebec, Canada). Mycophenolic acid was measured using a commercially available EMIT (Novartis Pharmaceuticals, Canada) assay. As the majority of patients were treated with low doses of cyclosporine after adding MMF, the area under the concentration-time curve (AUC) for cyclosporine showed a wide scatter ranging from 296 to 6400 ng x h/mL. The mean cyclosporine dose was 100 +/- 76 mg/m2 per day (range: 28 to 331). There was no correlation between MPA AUC and MPA dose, and there was substantial interindividual variation. However, there was a significant negative correlation between dose-normalized MPA AUC and cyclosporine AUC ( r2 = 0.23, p < 0.0220). When dividing the MPA profiles into two groups (11 and 12 patients) with a CyA AUC less than or greater than 1600 ng x h/mL, there was a significantly higher 8-hour concentration in the patients with the lower CyA AUC, secondary to a higher second peak. The data demonstrate that the cyclosporine AUC is a determining factor for the MPA AUC and that MPA dose should be reduced when cyclosporine dose is reduced to achieve the same MPA AUC. The significantly higher peak in the group with a lower CyA profile supports the concept of a dose-dependent cyclosporine-induced inhibition of MPA glucuronidation.

Genetic and amino acid analysis of the GL protein of Canadian, American and European equine arteritis virus isolates.

The genetic variation in equine arteritis virus (EAV) GL protein encoding gene was investigated. Nucleic and deduced amino acid sequences from 7 different EAV isolates, including 4 eastern Canadian field isolates, were compared with those of the Bucyrus reference strain. Nucleotide sequence identities between these isolates and the Bucyrus reference strain ranged from 87.5% (Vienna isolate) to 93.9% (11958 isolate). Amino acid identities with the Bucyrus reference strain varied from 90.2% (Vienna isolate) to 95.1% (19933 isolate). A 2nd potential N-linked glycosylation site was found at position 81 in the GL protein of all EAV isolates. Three amino acid substitutions at residue position 90 (Glu-->Val), position 101 (Ala-->Val or Thr), and position 119 (Val-->Leu, Phe or Ser) were also found in all EAV isolates. Phylogenetic analysis showed that the North American EAV isolates, including the Canadian isolates, and the European prototype Vienna isolate could be classified in 2 distinct groups. Three putative sequential antigenic sites were predicted in EAV GL protein. The 1st antigenic site (TAQRFT) was located at positions 24 to 29, and the 2nd antigenic site (RYDEHTA) at positions 47 to 53. The 3rd antigenic site was predicted to be located at positions 78 to 84 and showed the less conserved amino acid sequence.

Age-specific distribution of plasma amino acid concentrations in a healthy pediatric population.

Reference values were determined for 23 plasma free amino acids from measurements done in 148 healthy children ranging from 0 to 18 years of age. Amino acid analysis was performed by ion-exchange chromatography. We propose a graphic form of presenting the age-specific distribution of plasma amino acid concentrations where the 10th, 50th, and 90th quantiles are illustrated. Although each amino acid possesses its own pattern of distribution, we can identify five different profiles. Nine amino acids (alanine, arginine, asparagine, methionine, ornithine, phenylalanine, proline, threonine, and tyrosine) demonstrate a decrease in their concentrations during the first year of life; their concentrations then tend to increase throughout childhood and adolescence. Nine others (cystine, glutamine, glycine, histidine, isoleucine, leucine, lysine, tryptophan, and valine) show a steady increase throughout infancy, childhood, and adolescence. Five amino acids (aspartic acid, citrulline, glutamic acid, serine, and taurine) do not follow these two common profiles. For the first time, quantile curves are produced to illustrate the age-dependent variation of amino acid concentrations from infancy to adulthood. This alternative way of presenting amino acid concentrations may facilitate the follow-up of patients with inborn errors of amino acid metabolism.

Comparison of nucleic and amino acid sequences and phylogenetic analysis of the Gs protein of various equine arteritis virus isolates.

The genetic variation in equine arteritis virus (EAV) Gs protein encoding gene was investigated. Nucleic and deduced amino acid sequences from eight different EAV isolates (one European, two American and five Canadian isolates) were compared with those of the Bucyrus reference strain. Nucleotide and amino acid identities between these isolates and the Bucyrus reference strain ranged from 92.3 to 96.4%, and 93.2 to 95.5%, respectively. However, phylogenetic tree analysis and estimation of genetic distances based on the Gs protein encoding gene sequences showed that the European prototype Vienna strain, the American 87AR-A1 isolate and all other North American EAV isolates could be classified into three genetically divergent groups. Our results showed that the Gs protein-encoding gene can be subjected on the basis of phylogenetic analysis to genetic variation, as previously shown for the other three EAV structural protein (M, N and GL)-encoding genes.

Purification of lysophospholipase of human spermatozoa and its implication in the acrosome reaction.

The lysophospholipase of human spermatozoa was purified to homogeneity by sequential ion-exchange, gel filtration, and hydrophobic chromatography. The final preparation exhibited a single protein band on SDS-PAGE. The molecular mass of the enzyme was estimated to be 51 kDa by SDS-PAGE and 52 kDa by gel filtration. The optimal pH of this enzyme is 8.0. Polyclonal antibodies against lysophospholipase were prepared by placing the enzyme adsorbed on nitrocellulose directly into the spleen of rabbits. These antibodies were purified by protein A-agarose and by affigel-lysophospholipase chromatography. The purified antibodies and enzyme were used to study the possible role of lysophospholipase in the acrosome reaction. The addition of these antibodies led to an increase in the acrosome reaction, thus suggesting that inhibition of lysophospholipase produces a higher lysophosphatidylcholine concentration and results in an acrosome reaction level similar to that obtained by the calcium ionophore A23187. Immunofluorescence localization of the enzyme indicated that the enzyme is located on the head of spermatozoa. The purified sperm lysophospholipase and its specific antibodies represent important tools for the study of the regulation of this enzyme in reproductive processes. Furthermore, the study of this enzyme will allow evaluation of the mechanisms underlying the acrosome reaction.

Distribution of lysophospholipids and metabolism of platelet-activating factor in human follicular and peritoneal fluids.

The distribution of lysophosphatidylcholine, lyso-platelet-activating factor and platelet-activating factor (PAF) was studied in human plasma and in follicular and peritoneal fluid. In plasma, peritoneal and follicular fluids, 51%, 87% and 89%, respectively, of the total lipids were found in the protein fraction (the density > 1.21 fraction). Two forms of lysophospholipids were identified in this fraction: one of high affinity and one of low affinity for albumin. The metabolism of PAF in human follicular fluid, peritoneal fluid and plasma was also investigated. PAF-acetylhydrolase activity was found in both peritoneal and follicular fluids which induced a time-dependent hydrolysis of [3H]PAF. The half-life of PAF was estimated to be 7-12 min in plasma, 15-25 min in peritoneal fluid and approximately 2 h in follicular fluid. PAF-acetylhydrolase activity in embryo culture media supplemented with 10% serum was markedly inhibited by addition of commercial serum albumin. When 25 g albumin l-1 was added, 22% of [3H]PAF was hydrolysed h-1 compared with 72% in media without albumin. The concentrations of lysophosphatidylcholine measured in plasma, in follicular and peritoneal fluids were 252, 286 and 53 mumol l-1, respectively. The distribution of these lysophospholipids and the metabolism of PAF in the female genital tract fluids reported in the present study provide evidence for the involvement of these biologically active lipid mediators in a variety of reproductive processes including sperm-egg interactions and embryonic development.

Interference of lysophosphatidylcholine in hormone radioimmunoassays.

The interference of synthetic and naturally occurring detergents in immunoassays is well documented. In the present study, we evaluated the effect of lysophosphatidylcholine (LPC) and found that the lysophospholipid interfered with formation of the antigen-antibody complex in hormone immunoassays. In the presence of LPC (100 mumol/L), progesterone was overestimated by 29%. Furthermore, physiological concentrations of LPC (140 mumol/L) interfered with the assays of cortisol, progesterone, and aldosterone, resulting in overestimations of 35%, 30%, and 27%, respectively. The addition of albumin decreased the interference by LPC to 7% in the assay of cortisol and progesterone when the LPC:albumin ratio was unity. Adding cholesterol (100 mumol/L) also reduced by 50% the interference induced by LPC. Finally, treating plasma to increase the endogenous LPC concentration also resulted in interference in the cortisol assay. Thus, interpretation of the results of these assays should take into consideration the endogenous serum albumin:LPC ratio.

Platelet-activating factor acetylhydrolase activity in peritoneal fluids of women with endometriosis.

OBJECTIVE: To investigate whether platelet-activating factor acetylhydrolase may play a role in the pathophysiology of endometriosis. METHODS: Peritoneal fluids were obtained at laparoscopy from eight fertile women without endometriosis and 12 women with mild endometriosis. Platelet-activating factor acetylhydrolase activity was assayed in serum and peritoneal fluid by the release of 3H-acetate from [3H]-platelet-activating factor during a 15-minute incubation. RESULTS: Platelet-activating factor acetylhydrolase activity (mean +/- standard error of the mean) was significantly lower (P < .01) in peritoneal fluids from women with mild endometriosis (29.5 +/- 2.7%) than in controls (39.9 +/- 3.4%). The platelet-activating factor acetylhydrolase-cholesterol and -albumin ratios were also significantly lower in peritoneal fluids of women with endometriosis compared with controls (P < .006 and P < .012, respectively). CONCLUSION: These data suggest that mild endometriosis is associated with decreased platelet-activating factor acetylhydrolase activity in peritoneal fluid.