Genome characterization of lipid-containing marine bacteriophage PM2 by transposon insertion mutagenesis

Bacteriophage PM2 presently is the only member of the Corticoviridae family. The virion consists of a protein-rich lipid vesicle, which is surrounded by an icosahedral protein capsid. The lipid vesicle encloses a supercoiled circular double-stranded DNA genome of 10,079 bp. PM2 belongs to the marine phage community and is known to infect two gram-negative Pseudoalteromonas species. In this study, we present a characterization of the PM2 genome made using the in vitro transposon insertion mutagenesis approach. Analysis of 101 insertion mutants yielded information on the essential and dispensable regions of the PM2 genome and led to the identification of several new genes. A number of lysis-deficient mutants as well as mutants displaying delayed- and/or incomplete-lysis phenotypes were identified. This enabled us to identify novel lysis-associated genes with no resemblance to those previously described from other bacteriophage systems. Nonessential genome regions are discussed in the context of PM2 genome evolution.     

Decomposition rate of organic substrates in relation to the species diversity of soil saprophytic fungi

Despite the great interest concerning the relationship between species diversity and ecosystem functioning, there is virtually no knowledge as to how the diversity of decomposer microbes influences the decomposition rate of soil organic matter. We established a microcosm study in which the number of soil fungi was investigated in relation to the systems ability to (i) degrade raw coniferous forest humus, and (ii) use resources that were either added to the systems or released into the soils after a disturbance (drought). With the exception of the most diverse treatment, in each of the six replicates of each of the six diversity treatments (1, 3, 6, 12, 24 or 43 taxa), fungal taxa were randomly chosen from a pool of 43 commonly isolated fungal species of raw humus. Two months after initiation of the study CO₂ production increased as fungal diversity increased, but in the species-poor end of the diversity gradient only. Addition of various energy resources to the microcosms generally increased the level of soil respiration but did not affect the shape of the diversity-CO₂-production curve. Rewetting the soil after severe drought resulted in a rapid flush of CO₂, particularly in the most diverse communities. The biomass of the fungi in the non-disturbed soils, and soil NH₄-N concentration and soil pH in both disturbed and non-disturbed systems were slightly but significantly higher in the diverse than in the simple systems. Fungal species richness had no influence on the organic matter content of the humus at the end of the experiment. The results suggest that the functional efficiency of fungal communities can increase with the number of fungal taxa. This diversity effect was, however, significant at the species-poor end of the diversity gradient only, which implies considerable functional equivalency (redundancy) among the decomposer fungi.     

Direct in vivo monitoring of acute allergic reactions in human conjunctiva

Immediate allergic reactions are initiated by allergen-induced, specific IgE-mediated mast cell degranulation and involve leukocyte recruitment into the inflamed site. We compared conjunctival signs, symptoms, and in vivo leukocyte rolling and extravasation into sites of inflammation in five patients allergic to birch pollen and in 10 nonallergic controls who received a challenge to birch allergen or histamine. Both the specific allergen in allergic patients and histamine, both in patients and in healthy controls, induced symptoms and signs of an immediate allergic reaction together with leukocyte rolling within the conjunctival blood vessels. However, only allergen, not histamine, caused leukocyte extravasation into the site of inflammation in the allergic patients. Allergen also increased expression of endothelial P-selectin in conjunctival vessels and slowed the rolling of leukocytes which is required for their extravasation from blood circulation into the target tissue. Finally, i.v. heparin strongly reduced the number of slowly rolling cells during allergen- or histamine-induced reactions and this can probably hinder the leukocyte extravasation after allergen exposure. These findings suggest that slow rolling is required for leukocyte extravasation in acute allergic reactions, and it can be inhibited by heparin in vivo in therapeutically relevant conditions.     

Meta-analysis in assessment of the prevalence and annual incidence of Giardia spp. and Cryptosporidium spp. infections in humans in the Nordic countries

We aimed to apply the meta-analysis in the studies of protozoan pathogens in order to obtain a general overview of the prevalence and annual incidence of Giardia spp. and Cryptosporidium spp. infections in asymptomatic and symptomatic human populations in the Nordic countries of Denmark, Finland, Norway and Sweden. In combining the data of 13 clinically and methodologically non-heterogeneous studies published before 2004 using the random effects model with DerSimonian-Laird estimator, we estimated the prevalence (% prevalence: 95% confidence limits) of Giardia cases in the asymptomatic (i.e. no gastroenteric symptoms) general population to be 2.97% (2.64; 3.31) and in the symptomatic population 5.81% (5.34; 6.30). For Cryptosporidium the prevalences were 0.99% (0.81; 1.19) and 2.91% (2.71; 3.12), respectively. In analyzing the data, we estimated that there will be 4670 (4300; 5060) symptomatic cases of Giardia and 3340 (3110; 3580) symptomatic cases of Cryptosporidium annually per 100,000 general population in the Nordic countries. The vast majority of cases will remain unregistered in the national registers of infectious diseases, since for single registered cases there will be 254-867 cases of Giardia undetected/unregistered and 4072 to 15,181 cases of Cryptosporidium, respectively.     

Sex difference in the regulation of plasma high density lipoprotein cholesterol by genetic and environmental factors

Association between high density lipoprotein (HDL) cholesterol concentration and restriction fragment length polymorphisms at the cholesteryl ester transfer protein (CETP) gene locus was studied in a random population-based cohort of 526 Caucasian subjects (259 men, mean age 50.9 years, and 267 women, mean age 51.8 years). HDL cholesterol concentration was adjusted for age, body mass index, alcohol consumption, smoking and plasma triglyceride and low density lipoprotein cholesterol levels. In females, the HDL cholesterol levels were associated withTagIB polymorphism (1.46 mmol/1 in the B1B1 genotype, 1.56 mmol/l in B 1B2 and 1.72 mmol/1 in B2B2,P = 0.0001 for the trend). In contrast, this was not observed in men (1.24, 1.20, 1.27 mmol/l, NS). The association was seen even in women who were current smokers (1.41, 1.56, 1.75 mmol/l,n = 72,P = 0.007), but not in male smokers (1.26, 1.19, 1.14 mmol/l,n = 102, NS). In male non-smokers the association was weak (1.22, 1.20, 1.32 mmol/l,n = 157,P = 0.05). In postmenopausal women not receiving hormone replacement therapy (n = 108), the association continued to be present, although weaker (1.50, 1.58, 1.70 mmol/l,P = 0.06). CETP activity (n = 101) tended to be lower in subjects with the 132132 genotype. In conclusion, a clear-cut sex difference was observed in the genotype effect on plasma HDL cholesterol levels. The slight attenuation of the gene dosage effect after menopause suggests that the gender difference may be, at least in part, due to sex hormones. A genetic subgroup (men with the 132132 genotype) particularly susceptible to the HDL cholesterol decreasing effect of smoking could be demonstrated.Part of this work was presented at the 67th Scientific Sessions of the American Heart Association, Dallas, Texas, USA, 14–17 November, 1994     

Maturation of urinary proteoglycan excretion

Urinary proteoglycan excretion was studied using two newly established methods in subjects aged between 1 and 22 years. Analysis of glycan moieties showed an age-dependent decrease from 9.1±5.5 (SD) g/mol creatinine (n=5) at the age 1–6 years to 1.9±1.3 (n=5,P<0.01) in those aged 16–22 years. Marked qualitative changes in the proteoglycan electrophoretic pattern occurred during the first and second years of life. Two major proteoglycan bands with a molecular weight of 50 kDa decreased in intensity so that the pattern resembled the adult configuration after 6 years of age. The latter consisted of a major band with a molecular weight of 80–100 kDa, the bands corresponding to a molecular weight of 50 kDa and lighter bands of molecular weight around 32 kDa. These changes may be related to functional maturation of the kidney as a whole and to an increase in the number of mature nephrons.     

Palaeolimnological study of an environmental monitoring area,or, are there pristine lakes in Finland?

A palaeolimnological study of Lake Iso-Hietajärvi in Patvinsuo National Park, Lieksa, East Finland was conducted. The drainage area of the lake is one of four Integrated Monitoring areas established in Finland. Lead-210 dating reveals a period of increased sedimentation in the lake from 1920 to 1950. Increased atmospheric burden of several heavy metals in the order Pb > Cu> Zn > Ti = Al > Cr = Ni = V is recorded. The first to expand during the 19th century is Pb, whilst V increases after 1950. Sedimentary chlorophyll derivatives expand in the early part of the 20th century. Assemblages of diatoms and Cladocera were also changed somewhat during this time, but water quality seems not to have varied much: e.g. the diatom-inferred pH has remained in the range 6.4–6.8 (with a slight decrease) throughout the period of study.     

Occurrence of disialosyl groups in glycoproteins

The occurence of disialosyl (α-N-acetylneuraminyl-(2→8)-N-acetylneuraminyl) groups in glycoproteins was studied. It was found by methylation analysis that 8.5 % of N-acetylneuraminic acid in brain glycoproteins was substituted at C-8. The corresponding value was 16.6 % in brain gangliosides. The substituent was identified as neuraminic acid from its lability to neuraminidase treatment. The results demonstrate that not only gangliosides contain disialosyl groups, but these are also found in glycoproteins. The group is present in several types of carbohydrate units, with the highest proportion in N-glycosidic chains of large molecular size.     

Gene expression profiles in chondrosarcoma cells subjected to cyclic stretching and hydrostatic pressure. A cDNA array study

Mechanical forces have a profound effect on cartilage tissue and chondrocyte metabolism. Strenuous loading inhibits the cellular metabolism, while optimal level of loading at correct frequency raises an anabolic response in chondrocytes. In this study, we used Atlas Human Cancer cDNA array to investigate mRNA expression profiles in human chondrosarcoma cells stretched 8% for 6 hours at a frequency of 0.5 Hz. In addition, cultures were exposed to continuous and cyclic (0.5 Hz) 5 MPa hydrostatic pressure. Cyclic stretch had a more profound effect on the gene expression profiles than 5 MPa hydrostatic pressure. Several genes involved with the regulation of cell cycle were increased in stretched cells, as well as mRNAs for PDGF-B, glucose-1-phosphate uridylyltransferase, Tiam1, cdc37 homolog, Gem, integrin alpha6, and matrix metalloproteinase-3. Among down-regulated genes were plakoglobin, TGF-alpha, retinoic acid receptor-alpha and Wnt8b. A smaller number of changes was detected after pressure treatments. Plakoglobin was increased under cyclic and continuous 5 MPa hydrostatic pressure, while mitogen-activated protein kinase-9, proliferating cell nuclear antigen, Rad6, CD9 antigen, integrins alphaE and beta8, and vimentin were decreased. Cyclic and continuous pressurization induces a number of specific changes. In conclusion, a different set of genes were affected by three different types of mechanical stimuli applied on chondrosarcoma cells.