Clinical course of homozygous familial hypercholesterolemia during childhood: report on 4 unrelated patients with homozygous or compound heterozygous mutations in theLDLR gene

Natural history of the disease in 4 unrelated Polish children with homozygous familial hypercholesterolemia (FH) is described. Their phenotypic homozygosity was established by identification of known LDLR gene mutations on both alleles, respectively: p.G592E & p.G592E in Patient 1; p.G592E & p.C667Y in Patient 2; p.S177L & p.R350X in Patient 3; and p.G592E & deletion in the promoter region, exons 1 and 2 in Patient 4. Heterozygosity of the mutations was revealed in all patients' mothers and fathers (obligatory heterozygotes) and in 1 out of 4 siblings studied. FH was diagnosed at the age of 4 months to 9 years by cholesterol screening among family members of patients with early cardiovascular disease episodes. At the time of FH detection, the children were asymptomatic. Only in 2, some skin eruptions were found. Antihyperlipidemic therapy was started, including a lipid-lowering diet, cholestyramine, and HMG-CoA inhibitors if necessary. No cardiovascular symptoms appeared during the observation up to the age of 18, 20, 19, and 17 years, respectively. An increase in external carotid artery diameter was found in a patient at the age of 9 years, and LDL-apheresis was introduced in his therapy. We conclude that the analysis of LDLR gene mutations in the studied FH children made it possible to identify 4 presymptomatic FH homozygotes and to introduce early appropriate treatment. Multicenter analysis of such persons would finally determine if the early lipid-lowering procedures can significantly reduce the risk of premature cardiovascular disease in homozygous FH.     

Calculation of lipophilicity for Pt(II) complexes: experimental comparison of several methods

Platinum containing compounds are promising antitumor agents, but must enter cells before reaching their main biological target, namely DNA. Their distribution within the body, and hence their activity is to a large extent determined by their lipophilicity, thus there is a strong interest to develop computational methods to predict this important property. This study analyses accuracy of five methods, namely ALOGPS, KOWWIN, CLOGP and two quantum chemical approaches, to predict octanol/water partition coefficients (log P) for sets of 43 and 12 Pt(II) complexes, collected from the literature and measured by the authors, respectively. All methods gave generally poor results with mean absolute error (MAE) of between 0.8 and 3 log units for prediction of new compounds. Extension of the ALOGPS program with data from the literature set resulted in the best prediction ability, MAE = 0.46, for the measured molecules. The program was also able to correctly predict errors in calculated log P values. It is freely available for interactive use at http://www.vcclab.org.     

Model-based inference of neutralizing antibody avidities against influenza virus

To assess the response to vaccination, quantity (concentration) and quality (avidity) of neutralizing antibodies are the most important parameters. Specifically, an increase in avidity indicates germinal center formation, which is required for establishing long-term protection. For influenza, the classical hemagglutination inhibition (HI) assay, however, quantifies a combination of both, and to separately determine avidity requires high experimental effort. We developed from first principles a biophysical model of hemagglutination inhibition to infer IgG antibody avidities from measured HI titers and IgG concentrations. The model accurately describes the relationship between neutralizing antibody concentration/avidity and HI titer, and explains quantitative aspects of the HI assay, such as robustness to pipetting errors and detection limit. We applied our model to infer avidities against the pandemic 2009 H1N1 influenza virus in vaccinated patients (n = 45) after hematopoietic stem cell transplantation (HSCT) and validated our results with independent avidity measurements using an enzyme-linked immunosorbent assay with urea elution. Avidities inferred by the model correlated with experimentally determined avidities (ρ = 0.54, 95% CI = [0.31, 0.70], P < 10⁻⁴). The model predicted that increases in IgG concentration mainly contribute to the observed HI titer increases in HSCT patients and that immunosuppressive treatment is associated with lower baseline avidities. Since our approach requires only easy-to-establish measurements as input, we anticipate that it will help to disentangle causes for poor vaccination outcomes also in larger patient populations. This study demonstrates that biophysical modelling can provide quantitative insights into agglutination assays and complement experimental measurements to refine antibody response analyses.     

Occurrence and variability of River Habitat Survey features across Europe and the consequences for data collection and evaluation

River Habitat Survey (RHS) data collected for the EU-funded STAR project was used to identify hydromorphological characteristic features of rivers in four European regions namely: lowlands; mountain; the Alps; and the Mediterranean. Using RHS attributes, Habitat Quality Assessment (HQA) – a measure of natural habitat diversity, and Habitat Modification Score (HMS) – a measure of anthropogenic modification, we identified considerable differences in frequency, diversity and evenness of features between the regions. A relatively small subset of features clearly distinguish the hydromorphological characters of lowland, Alpine and southern European rivers. It was more difficult to distinguish mountain rivers from Alpine rivers. The highest statistical differences are observed between Lowland and Mountain region. Within the four regions studied the RHS attributes that most strongly influence the HQA and HMS indices were identified. We conclude that specific effort should be made to ensure these are recorded properly as part of the quality control of RHS data. Electronic supplementary material Electronic supplementary material is available for this article at and accessible for authorised users.     

Excretion of cytoplasmic proteins (ECP) in Staphylococcus aureus

E xcretion of c ytoplasmic p roteins (ECP) is a common physiological feature in bacteria and eukaryotes. However, how these proteins without a typical signal peptide are excreted in bacteria is poorly understood. We studied the excretion pattern of cytoplasmic proteins using two glycolytic model enzymes, aldolase and enolase, and show that their excretion takes place mainly during the exponential growth phase in Staphylococcus aureus very similar to that of Sbi, an IgG‐binding protein, which is secreted via the Sec‐pathway. The amount of excreted enolase is substantial and is comparable with that of Sbi. For localization of the exit site, we fused aldolase and enolase with the peptidoglycan‐binding motif, LysM, to trap the enzymes at the cell wall. With both immune fluorescence labeling and immunogold localization on electron microscopic thin sections aldolase and enolase were found apart from the cytoplasmic area particularly in the cross wall and at the septal cleft of dividing cells, whereas the non‐excreted Ndh2, a soluble NADH:quinone oxidoreductase, is only seen attached to the inner side of the cytoplasmic membrane. The selectivity, the timing and the localization suggest that ECP is not a result of unspecific cell lysis but is mediated by an as yet unknown mechanism.     

Experimentelle Untersuchungen Über die Einwirkung der Temperatur 0‡ C. auf die Entwicklung der Echinideneier

Ohne ZusammenfassungVorgelegt in der Sitzung der Akademie der Wissenschaften in Krakau vom 1. Juli 1912. Vorl. Mitteil. im Bull. de l'Acad. des Sc. de Cracovie. 1912.     

Estimating the 3D Pore Size Distribution of Biopolymer Networks from Directionally Biased Data

The pore size of biopolymer networks governs their mechanical properties and strongly impacts the behavior of embedded cells. Confocal reflection microscopy and second harmonic generation microscopy are widely used to image biopolymer networks; however, both techniques fail to resolve vertically oriented fibers. Here, we describe how such directionally biased data can be used to estimate the network pore size. We first determine the distribution of distances from random points in the fluid phase to the nearest fiber. This distribution follows a Rayleigh distribution, regardless of isotropy and data bias, and is fully described by a single parameter—the characteristic pore size of the network. The bias of the pore size estimate due to the missing fibers can be corrected by multiplication with the square root of the visible network fraction. We experimentally verify the validity of this approach by comparing our estimates with data obtained using confocal fluorescence microscopy, which represents the full structure of the network. As an important application, we investigate the pore size dependence of collagen and fibrin networks on protein concentration. We find that the pore size decreases with the square root of the concentration, consistent with a total fiber length that scales linearly with concentration.     

Use of benthic diatom communities to evaluate water quality in rivers of southern Poland

Biological and chemical data were processed to estimate trophic stage and degree of pollution in several streams and rivers in southern Poland. The majority were eutrophic and some of them heavily polluted; only a few were oligo-mesotrophic. The differences in the water quality of the rivers were reflected by different types of diatom community and also by the values for some diatom indices, which were calculated using the latest version of the 'Omnidia' database software. Except for the Sládeček's index, all diatom indices correlated significantly with organic load (COD), oxygen concentration, conductivity and most of the measured ions. Some indices showed a significant negative correlation with trophic level (expressed by NH₄-N and PO₄-P). In general, IPS (Specific Pollution Sensitivity Index) and GDI (Generic Diatom Index) indices gave the best results. Among the investigated diatom communities, only a few taxa indicated oligo-mesotrophy and oligo-β-mesosaprobity. Most of the sites were characterised by a greater relative contribution of eutraphent and tolerant ones as well as α-mesosaprobic and polysaprobic diatoms. This study suggests that the structure of benthic diatom communities and diatom indices, especially GDI, can be applied for monitoring rivers in Poland. This revised version was published online in June 2006 with corrections to the Cover Date.     

Serological relationship between three varieties of Bacillus popilliae

The serological relationship between three varieties of Bacillus popilliae was investigated by double diffusion and immunoelectrophoresis, using antigens prepared from homogenized vegetative cells in the logarithmic growth phase. The results showed the existence of a close serological relationship among the three varieties in agreement with a recently proposed taxonomic scheme for the milky disease bacteria (Wyss, 1971).