Serum pneumoproteins and biomarkers of exposure to urban air pollution: a cross-sectional comparison of policemen and foresters

Very few biomarkers are available for the non-invasive detection of effects of urban air pollution on the respiratory tract. The objective was to evaluate whether Clara cell protein (CC16) and surfactant-associated protein-A (SP-A), two pulmonary secretory proteins, were useful in the detection of effects of urban air pollutants on the pulmonary epithelium. These proteins were determined in the serum of 53 policemen working in Brussels, Belgium, and a control group of 59 foresters working in the countryside. Except for ozone (O₃), annual concentrations of the main air pollutants (PM₁₀, NO₂, CO, SO₂ and benzene) were significantly higher in Brussels than in the country. The proportion of smokers was lower in urban policemen compared with foresters, but they smoked on average a similar number of cigarettes per day as confirmed by their urinary excretion of cotinine. Muconic acid, a marker of benzene exposure, was significantly higher in urban policemen than in foresters, in both smokers and non-smokers. Multiple regression analysis showed that the type of work, smoking habits and time spent outdoors and in a car were significant determinants of benzene uptake. Tobacco smoking impaired lung function to a similar extent in urban policemen and foresters. The serum levels of SP-A were significantly increased in smokers but were not different between policemen and foresters. Serum CC16 was significantly reduced by tobacco smoking and slightly decreased in policemen compared with foresters. Interestingly, the reduction of serum CC16 was more pronounced in the subgroup of traffic compared with survey policemen, the latter being also less exposed to benzene. The results suggest that serum pneumoproteins and especially serum CC16 could be useful in the detection of chronic effects of urban air pollutants on the respiratory epithelium of populations particularly at risk.     

Adipose Angiotensinogen Secretion,Blood Pressure,and AGT M235T Polymorphism in Obese Patients

Objective: To investigate AGT secretion in cultured adipocytes from obese patients and its relationship with obesity‐related phenotypes, blood pressure, and the M235T polymorphism in the AGT gene. Research Methods and Procedures: Measurements, including anthropometry, body composition (DXA), and blood pressure, were performed in 61 overweight or obese women (BMI: 28 to 68 kg/m²). A subcutaneous abdominal adipose tissue biopsy was used for adipocyte size determination and quantification of AGT secretion in the medium of cultured adipocytes. AGT M235T genotype was determined using polymerase chain reaction‐restriction fragment length polymorphism. Results: Adipose secretion of the AGT protein (range, 140 to 2575 ng/10⁶ cells/24 h) was not significantly correlated with BMI, body fat, or blood pressure and did not vary according to the M235T polymorphism in the AGT gene. However, the AGT M235T polymorphism was associated with adipocyte size (111.6 ± 2.8, 108.8 ± 1.9, 118.2 ± 2.6 μm in MM, MT, and TT genotypes, respectively; p < 0.01) after adjustment for age and fat mass. An association between the AGT M235T polymorphism and adipocyte size (p < 0.02 adjusted for sex, age, and BMI) was found in another independent sample of 106 obese subjects (sex ratio, M/F 16/90; BMI, 29 to 70 kg/m²). Discussion: In cultured adipocytes from obese subjects, AGT secretion was not associated with body fat phenotypes, blood pressure, or fat cell size. However, results from two independent studies suggest an association between the AGT M235T polymorphism and adipocyte size.     

Control of DNA excision efficiency in Paramecium

Programmed excision of internal eliminated sequences (IESs) occurs at thousands of sites in ciliate genomes. How this is controlled is largely unknown. Here, we report the characterization of the non-efficiently excised 156ψG-11 IES from Paramecium primaurelia strain 156 and that of the efficiently excised 168ψG-11 IES, an allelic variant from strain 168. Then, we report a genetic and molecular analysis of IES excision efficiency in F₁ progeny derived from interstrain crosses and in F₂ homozygous progeny derived from F₁ autogamy. IES 168ψG-11 excision efficiency was ~100% in all cases. IES 156ψG-11 excision efficiency was 19 ± 13% in F₁ progeny and 0.6 ± 1.1% in F₂ progeny. No trans-excision event between IESs 156ψG-11 and 168ψG-11 was detected within the F₁ progeny. These data demonstrate that the excision efficiency of this IES is variable and controlled by a cis-acting element. This should encompass positions 8 and/or 9 of the right IES end, which display allele differences. Finally, the 30-fold stimulation of IES 156ψG-11 excision efficiency within F₁ progeny relative to F₂ progeny demonstrates that Paramecium IES excision efficiency is sensitive either to a conjugation-specific trans-acting factor provided by the zygotic genome, or to homologous chromosome cross-talk.     

Sensitive determination of vinorelbine and its metabolites in human serum using liquid chromatography-electrospray mass spectrometry

A liquid chromatography-electrospray mass spectrometry method was developed for the quantitation of vinorelbine (VNB) and two metabolites, vinorelbine N-oxide (VNO) and deacetyl vinorelbine (DAV) in human serum. The limits of quantitation (LOQ) reached 0.5 ng/ml for both VNB and VNO and 1 ng/ml for DAV. The method was proved linear in the range of LOQs up to 1000 ng/ml, and extraction recovery was 80% on average for the three compounds. It was applied to the pharmacokinetic monitoring of vinorelbine and, for the first time, to the detection of VNO in the serum of patients suffering from non-small-cell lung cancer.     

On-line identification of further flavone C- and O-glycosides from sugarcane (Saccharum officinarum L., Gramineae) by HPLC-UV-MS

HPLC-UV-MS (APCI-MS/MS and CID/MS) was utilised for the identification of eight additional flavone glycosides from sugarcane (Saccharum officinarum L., Gramineae) extracts of bagasse, leaves and juice ('garapa'). Relevant information about substitution patterns was obtained through UV detection using post-column addition of shift reagents, while tandem MS provided structural information confirming the proposed structures of the C-glycosides vitexin, orientin, luteolin-8-C-(rhamnosylglucoside), 4',5'-dimethyl-luteolin-8-C-glycoside and the isomeric pair schaftoside-isoschaftoside, besides the O-glycosides tricin-7-O-neohesperidoside and tricin-7-O-glycoside.     

Improved gene targeting in Magnaporthe grisea by inactivation of MgKU80 required for non-homologous end joining

The ascomycete Magnaporthe grisea is a model species for the study of plant fungal interactions. As in many filamentous fungi, targeted gene replacement occurs at low frequency in M. grisea (average 7%). mus52/KU80 is a gene essential for non-homologous end joining (NHEJ) of DNA double-strand breaks. Its deletion increases the frequency of targeted gene replacement in fungi [Ninomiya, Y., Suzuki, K., Ishii, C., Inoue, H., 2004. Highly efficient gene replacements in Neurospora strains deficient for non-homologous end joining. Proc. Natl. Acad. Sci. USA 101(33), 12248-53]. M. grisea KU80 deletion mutants were constructed and displayed wild-type phenotypes regarding pathogenicity, growth, sporulation and mating. MgADE4 targeted gene replacement frequency was increased in Deltaku80 mutants (80% vs 5%) and high frequencies (>80%) were observed at seven other loci. However, the deletion of MgKU80 did not increase the frequency of ACE1 replacement indicating that this locus has an intrinsic reduced ability for gene replacement. These results open the way to large-scale reverse genetics experiments in M. grisea facilitating the study of the infection process.     

Nesprin-3, a novel outer nuclear membrane protein, associates with the cytoskeletal linker protein plectin

Despite their importance in cell biology, the mechanisms that maintain the nucleus in its proper position in the cell are not well understood. This is primarily the result of an incomplete knowledge of the proteins in the outer nuclear membrane (ONM) that are able to associate with the different cytoskeletal systems. Two related ONM proteins, nuclear envelope spectrin repeat (nesprin)-1 and -2, are known to make direct connections with the actin cytoskeleton through their NH2-terminal actin-binding domain (ABD). We have now isolated a third member of the nesprin family that lacks an ABD and instead binds to the plakin family member plectin, which can associate with the intermediate filament (IF) system. Overexpression of nesprin-3 results in a dramatic recruitment of plectin to the nuclear perimeter, which is where these two molecules are colocalized with both keratin-6 and -14. Importantly, plectin binds to the integrin alpha6beta4 at the cell surface and to nesprin-3 at the ONM in keratinocytes, suggesting that there is a continuous connection between the nucleus and the extracellular matrix through the IF cytoskeleton.     

Antibacterial Peptides Are Present in Chromaffin Cell Secretory Granules

1. Antibacterial activity has recently been associated with the soluble matrix of bovine chromaffin granules. Furthermore, this activity was detected in the contents secreted from cultured chromaffin cells following stimulation.2. The agents responsible for the inhibition of Gram+ and Gram– bacteria growth are granular peptides acting in the micromolar range or below. In secretory granules, these peptides are generated from cleavage of chromogranins and proenkephalin A and are released together with catecholamines into the circulation.3. Secretolytin and enkelytin are the best characterized; these two peptides share sequence homology and similar antibacterial activity with insect cecropins and intestinal diazepam-binding inhibitor. For some of the peptides derived from chromogranin A, posttranslational modifications were essential since antibacterial activity was expressed only when peptides were phosphorylated and/or glycosylated.4. The significance of this activity is not yet understood. It may be reminiscent of some primitive defense mechanism or may serve as a first barrier to bacteria infection during stress, as these peptides are secreted along with catecholamines.     

Methicillin-resistant staphylococcal colonization in clinically normal dogs and horses in the community

AIMS: To evaluate the prevalence of methicillin-resistant staphylococcal (MRS) colonization in clinically normal dogs and horses in the community. METHODS AND RESULTS: Three hundred clinically normal horses and 200 clinically normal dogs were enrolled. One nasal swab was collected from each horse. Two swabs were taken from each dog: (i) from an anterior nare, and (ii) a combination of the perineal area and 0.5 cm into the anus. Enrichment cultures were performed. Methicillin-resistant Staphylococcus aureus (MRSA) was not identified. Methicillin-resistant Staphylococcus intermedius (MRSI) was isolated from the nasal swab from three dogs. Methicillin-resistant coagulase negative staphylococci (MRCoNS) were isolated from 126/300 (42%) horses and 26/200 (13%) dogs. CONCLUSIONS: At present MRSI is not considered to be a significant zoonotic concern; however, it may become an important pathogen in dogs. MRCoNS mostly cause disease in compromised human or animal hosts. However, these bacteria can serve as reservoirs of resistance determinants in the community, which could lead to the emergence of novel MRSA strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of the prevalence of MRS colonization in clinically normal dogs in a community setting. Continued surveillance is indicated to determine whether MRSA will emerge in the animal population and become a concern for animal disease and zoonotic infection.