Search

Boreal forests contain diverse fungal communities that form essential ectomycorrhizal symbioses with trees. To determine the effects of lead (Pb) contamination on ectomycorrhizal fungal communities associated with the dominant pine (Pinus sylvestris L.), we surveyed sporocarps for 3 years, analyzed morphotyped ectomycorrhizal root tips by direct sequencing, and 454-sequenced fungal communities that grew into in-growth bags during a 2-year incubation at a shooting range where sectors vary in the Pb load. We recorded a total of 32 ectomycorrhizal fungi that formed conspicuous sporocarps, 27 ectomycorrhizal fungal phylotypes from 294 root tips, and 116 ectomycorrhizal fungal operation taxonomic unit (OTUs) from a total of 8194 internal transcribed spacer-2 454 sequences. Our ordination analyses by nonparametric multidimensional scaling (NMS) indicated that the Pb enrichment induced a shift in the ectomycorrhizal community composition. This was visible as indicative trends in the sporocarp and root tip data sets, but was explicitly clear in the communities observed in the in-growth bags. The compositional shift in the ectomycorrhizal community was mainly attributable to an increase in the frequencies of OTUs assigned to genus Thelephora and to a decrease in the OTUs assigned to Pseudotomentella, Suillus, and Tylospora in Pb-contaminated areas when compared with the control. While the compositional shifts are clear, their functional consequences for the dominant trees or soil ecosystem function remain undetermined., (© 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)

Aims: To study the ability of automated ribotyping to characterize Obesumbacterium proteus and Hafnia alvei, to design primers and to evaluate standard end-point and real-time PCR for the detection of O. proteus biotype 1 in beer and in brewers's yeast-containing samples., Methods and Results: Automated ribotyping was carried out using the standard method with EcoRI and PvuII. The digestions with both enzymes clearly differentiated O. proteus biotypes 1 and 2 and H. alvei. PCR primers were designed according to the 16S rRNA gene sequence of the O. proteus type strain. Two primer sets (Obs137-Obs558 and Obs137-Obs617) detected O. proteus biotype 1 and H. alvei but not O. proteus biotype 2 or other tested beer spoilage bacteria (40 species) in the end-point and real-time PCR, indicating their high specificity. The detection limit for O. proteus was 160-1600 CFU 100 ml(-1) beer in the end-point PCR reactions and < or =160 CFU 100 ml(-1) beer in the real-time PCR reactions. More cells (from 16 to 3200) were needed for detection in the presence of brewer's yeast cells., Conclusions: Automated ribotyping is a useful tool to characterize and identify O. proteus and H. alvei isolates. The designed primers are suitable for the rapid detection of O. proteus biotype 1 and H. alvei in brewery samples by PCR., Significance and Impact of the Study: Automated ribotyping and PCR could improve microbiological quality control in breweries by facilitating the detection, identification and tracing of spoilage bacteria.

Apolipoprotein (apo)E gene epsilon4 allele carrier status modulates the responses of lipoprotein metabolism to hormone-replacement therapy (HRT). We investigated the effect of long-term HRT on the progression of atherosclerosis in postmenopausal women with or without apoE epsilon4 allele. One hundred forty-one nonsmoking postmenopausal women, 45-71 yr old, were divided into 3 groups based on the use of HRT. The HRT-EVP group (n = 61) used sequential estradiol valerate (EV) plus progestin (P), the HRT-EV group used EV alone (n = 40), and a control group had no HRT. Of these 141 women, 93 participated in a 5-yr follow-up study in 1998. In addition to serum lipid concentration and apoE genotype, the atherosclerosis severity score of the abdominal aorta and carotid arteries was determined by sonography. In apoE4-negative subjects, the progression of atherosclerosis severity score was significantly faster in control than in the HRT groups (genotype-by-time interaction P = 0.0026); whereas in apoE4-positive subjects, there were no significant differences in atherosclerosis severity score progression between the control and HRT groups. The effects of HRT on atherosclerosis progression in subjects with no apo epsilon4-allele seems to be especially beneficial, compared with controls with same phenotype status but without HRT. These results may help us to understand, in more detail, the benefit and possible risk of HRT on atherosclerotic diseases.

We investigated the means by which drug resistance emerges among drug-susceptible Mycobacterium tuberculosis strains during antituberculosis therapy. Patients who experienced failure of treatment for active pulmonary tuberculosis, who initially received diagnoses of infection with drug-susceptible M. tuberculosis, and who had had at least 3 isolates tested for drug susceptibility were selected from a 6-year period in the Estonian National Reference Laboratory archive. Eleven patients from whom 35 sequential isolates of M. tuberculosis had been obtained were recruited into the study. Their clinical data and treatment charts were analyzed and correlated with drug-susceptibility patterns and IS6110 restriction fragment-length polymorphism (RFLP) profiles. Six patients excreted isogenic drug-susceptible M. tuberculosis strains, whereas, in the other 5 patients, the isolated strain shifted from a susceptible to a resistant phenotype. In all cases, this shift correlated to a shift in RFLP pattern, which showed reinfection with a new strain. Exogenous reinfection with drug-resistant M. tuberculosis may be misinterpreted as the emergence of drug resistance if molecular testing techniques are not used.

The tolerance to, and degradation of m-toluate by Scots pine (Pinus sylvestris), a symbiotic mycorrhizal fungus (Suillus bovinus) and Pseudomonas fluorescens strains, with or without m-toluate-degrading capacity, was determined individually and in all symbiotic/associative plant-microbe combinations. Fungal survival on medium with m-toluate was increased in co-culture with the degradative bacterial strains on agar plates (up to 0.02%, w/v). When fungi were grown in mycorrhizal association with Scots pine seedlings in test-tube microcosms containing expanded clay pellets and growth media, the fungus was able to withstand m-toluate concentrations up to 2.0%, w/v in all treatments. The seedling tolerance remained unaltered regardless of the presence or absence of mycorrhizal fungi or biodegradative bacteria. Reduction in m-toluate levels was only detected in treatments inoculated with bacterial strains harbouring TOL catabolic plasmids. The plant and fungus, alone or in mycorrhizal symbiosis, were unable to cleave m-toluate. The presence of easily available plant-derived carbon sources did not impede m-toluate degradation by the bacteria in the mycorrhizosphere.

Background: The present study was undertaken to clarify the progression of urinary albumin excretion rate (UAER) in non-insulin-dependent diabetic (NIDD) patients 6 years after diagnosis, and to elucidate the risk factors of nephropathy., Methods: This is a population-based controlled (baseline) cohort study. The prospective evaluation utilized the diabetic patients as internal controls. The setting was an urban primary health care centre. Main outcome measures were the UAER-24 h and fractional urinary albumin excretion rate (FAC) and their relation to mean blood pressure, haemoglobin Alc, fasting serum insulin and cholesterol and renal size., Results: UAER (mg/24 h) was increased (geometric mean, quartile 1 and 3) in the diabetic patients at baseline, compared to the non-diabetic control subjects; 21 (10 and 33) versus 12 (8 and 15), P = 0.0001 (Wilcoxon's rank test). The UAER-24 h was not increased in diabetic subjects at follow-up; 24 (7 and 49) P = 0.3791 versus diabetic subjects at baseline. Eighteen per cent of normoalbuminuric (UAER < 30mg/24 h) patients developed microalbuminuria (UAER = 30-300 mg/24 h) and 3% clinical nephropathy (UAER > 300 mg/24 h). Of the microalbuminuric subjects 19% progressed to clinical nephropathy, 46% remained microalbuminuric and 35% remitted to normoalbuminuria. Serum insulin concentration, after assessment of confounding factors, measured at the baseline predicted the UAER for all diabetic subjects at follow-up in multiple linear regression analysis in an independent and significant way (P = 0.01). Serum insulin concentration (P = 0.034) and diuretic therapy (P = 0.050) at baseline independently predicted the outcome of the categorical variable progressor/nonprogressor (n = 22/86) based on the UAER-24 h at baseline and at follow-up., Conclusions: Progression of the UAER during the first 6 years is found among approximately every fifth NIDD subject who develops either microalbuminuria (from normoalbuminuria) or clinical nephropathy (from microalbuminuria). The role of serum insulin (insulin resistance) or some factor associated with it, is suggestive in the genesis of kidney disease.

In total, 814 patients with clinically suspected tuberculosis were examined at the Raoul Follerau Hospital in Bissau, Guinea-Bissau. Sputum samples were collected, and cultures were grown on Löwenstein-Jensen medium. Isolates were further characterized by standard biochemical methods and nucleic acid probes for Mycobacterium tuberculosis and Mycobacterium avium complex (MAC). Serum samples were collected and analyzed for antibodies against human immunodeficiency virus (HIV) types 1 and 2. Of 17 patients who had MAC organisms in sputum, 2 were HIV-2-positive and none was HIV-1-positive. Of the total 814 patients, 189 had culture-verified tuberculosis; 36 (19%) of them were HIV-2-positive. Thus, of 206 patients with sputum culture positive for M. tuberculosis or MAC, 17 (8%) had MAC organisms in sputum. MAC infection may be the cause of some treatment failures in areas where MAC pulmonary infection is common.

We utilized the databases of a hospital information system to select for determination of reference values various individual hospitalized patients on the basis of their diagnoses at discharge. The nonparametric 2.5-97.5% "health-related" reference intervals were calculated for hemoglobin concentration, mean corpuscular volume (MCV), and erythrocyte count for both sexes. After excluding patients with diseases possibly affecting erythrocyte variables, we obtained a final group of 1786 women and 1450 men, ages 20-65 years, who were studied in age groups of 20-30, 30-45, 45-55, and 55-65 years. The upper reference limits of the MCV results obtained from hospitalized patients were higher than those produced conventionally from healthy individuals, as would be intuitively suggested by clinical experience. This method, based on selection by diagnosis, could be applicable to various analytes measured in hospital laboratories, provided sufficient data are available as databases.

Background: Mutations in the p53 gene resulting in the accumulation of altered p53 proteins with prolonged half-life have been found in a large variety of human malignancies., Purpose: We studied the significance of p53 protein accumulation in prostatic carcinoma., Methods: The material consisted of 137 paraffin-embedded, primary prostatic carcinomas. Accumulation of p53 protein was studied by immunohistochemical staining using a polyclonal p53-specific CM-1 antibody. Proliferation activity was determined by DNA flow cytometry and by immunohistochemical detection of proliferative cell nuclear antigen (PCNA) using a monoclonal PC10 antibody., Results: Eight (6%) of the tumors showed intense p53 staining in more than 20% of the tumor cells, 15 (11%) had only lower level immunoreactivity, and 114 (83%) showed no staining. High-level p53 accumulation was associated with high histologic grade (P less than .001), DNA aneuploidy (P less than .05), and high cell proliferation rate as defined by flow cytometric S-phase analysis (P less than .01) or PCNA expression (P less than .01). High-level p53 accumulation predicted short, progression-free interval (P less than .01) and poor survival (P less than .001), with about a 12-fold relative risk of death as compared with p53-negative cases. Low-level p53 accumulation had no prognostic significance., Conclusions: Accumulation of p53 confers proliferative advantage for prostatic carcinoma cells and defines a small subgroup of highly malignant carcinomas.

A new biological alcohol marker, the ratio between the acetaldehyde-induced haemoglobin fraction HbA1ach and the glycated haemoglobin fraction HbA1c (HbA1ach/HbA1c), was studied in association with data obtained in a questionnaire, the Malmö modified Michigan alcoholism screening test (Mm-MAST), completed by 270 consecutive middle-aged men participating in a voluntary health screening. None of them had earlier alcoholic health or alcohol-dependent social problems. The nine-question Mm-MAST correlated positively with the ratio of HbA1ach/HbA1c (r = 0.272, P less than 0.001). In the three drinker groups defined according to the questionnaire, HbA1ach/HbA1c values were significantly higher among heavy drinkers (mean = 40, S.E.M. = 1) than among teetotallers (mean = 32, S.E.M. = 2, P less than 0.01) or social drinkers (mean = 35, S.E.M. = 1, P less than 0.01). In the groups formed according to the respondents' own alcohol anamnesis there was no significant difference between the marker values, but the Mm-MAST score was significantly higher in both social (P less than 0.001) and heavy drinkers' (P less than 0.001) groups as compared with teetotallers. This shows that the questionnaire detected heavy drinkers better than the self-given anamnesis. The correlation of HbA1ach/HbA1c and the questionnaire means that the biological marker studied represents a new possibility to identify heavy drinkers before alcoholic health problems or social consequences have appeared.

We studied the effect of one haemodialysis treatment on the plasma concentrations of intact parathyroid hormone (PTH), total immunoreactive PTH (determined with an antibody against the mid-molecular part of the hormone), and ionised calcium in 25 patients on maintenance dialysis for chronic renal failure. During dialysis the calcium concentration in the external fluid was 1.75 mmol/l, which led to an increase in the plasma ionised calcium concentration from 1.23 +/- 0.07 mmol/l (mean +/- SD) at commencement of dialysis to a final 1.35 +/- 0.07 mmol/l (P less than 0.001). The plasma concentration of intact PTH decreased from 27.4 +/- 26.3 pmol/l to 13.0 +/- 19.1 pmol/l (P less than 0.001) during the treatment. Total immunoreactive PTH did not change, reflecting poor dialysability of PTH and PTH fragments. We also compared the dialysis-induced changes in the plasma concentrations of intact PTH and ionised calcium with those induced by a calcium infusion test. We conclude that in the majority of dialysis patients, one haemodialysis treatment with a relatively high external fluid calcium concentration can be used to assess the suppressibility of secondary hyperparathyroidism.

In the present study it was found that uraemic plasma contains inhibitors which affect insulin binding on erythrocyte insulin receptors. Insulin radioreceptor assays performed in the absence of uraemic plasma revealed no defects in the receptors of the uraemic erythrocytes. Uraemic plasma caused a 48% decrease in insulin binding on healthy erythrocytes at a high (10.1 ng/ml) insulin concentration, but only a 17% decrease at a low (70 pg/ml) insulin concentration. The inhibition of insulin binding by uraemic plasma decreased during a haemodialysis treatment from 18% to 13% at the low insulin concentration, and from 30% to 23% at the high insulin concentration. Insulin insensitivity in uraemia appears to be due to inhibition of the insulin receptors.