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Mutations in the ABCC6 gene result in calcification diseases such as pseudoxanthoma elasticum or Generalized Arterial Calcification of Infancy. Generation of antibodies recognizing an extracellular (EC) epitope of ABCC6 has been hampered by the short EC segments of the protein. To overcome this limitation, we immunized bovine FcRn transgenic mice exhibiting an augmented humoral immune response with Human Embryonic Kidney 293 cells cells expressing human ABCC6 (hABCC6). We obtained a monoclonal antibody recognizing an EC epitope of hABCC6 that we named mEChC6. Limited proteolysis revealed that the epitope is within a loop in the N-terminal half of ABCC6 and probably spans amino acids 338-347. mEChC6 recognizes hABCC6 in the liver of hABCC6 transgenic mice, verifying both specificity and EC binding to intact hepatocytes.

Summary The prevention of enormous crop losses caused by pesticide‐resistant fungi is a serious challenge in agriculture. Application of alternative fungicides, such as antifungal proteins and peptides, provides a promising basis to overcome this problem; however, their direct use in fields suffers limitations, such as high cost of production, low stability, narrow antifungal spectrum and toxicity on plant or mammalian cells. Recently, we demonstrated that a Penicillium chrysogenum‐based expression system provides a feasible tool for economic production of P. chrysogenum antifungal protein (PAF) and a rational designed variant (PAFopt), in which the evolutionary conserved γ‐core motif was modified to increase antifungal activity. In the present study, we report for the first time that γ‐core modulation influences the antifungal spectrum and efficacy of PAF against important plant pathogenic ascomycetes, and the synthetic γ‐core peptide Pγopt, a derivative of PAFopt, is antifungal active against these pathogens in vitro. Finally, we proved the protective potential of PAF against Botrytis cinerea infection in tomato plant leaves. The lack of any toxic effects on mammalian cells and plant seedlings, as well as the high tolerance to harsh environmental conditions and proteolytic degradation further strengthen our concept for applicability of these proteins and peptide in agriculture., The emerging number of crop losses due to infection or contamination caused by pesticide‐resistant pre‐ and post‐harvest plant pathogenic fungi urges the need for the development of fundamentally new and safe antifungal strategies in the agriculture, and the cysteine‐rich, highly stable antifungal peptides and proteins from filamentous ascomycetes are promising candidates in this respect. We report for the first time that γ‐core modulation influences the antifungal spectrum and efficacy of Penicillium chrysogenum antifungal protein (PAF) against important plant pathogenic ascomycetes, and the synthetic γ‐core peptide Pγopt, a derivative of an engineered variant of PAF is antifungal active against these pathogens. We also prove the protective potential of PAF against Botrytis cinerea infection in tomato plant leaves, and the potential applicability of PAF, and its engineered variant as biofungicides in the agriculture as they do not show any toxic effects on mammalian cells and plant seedlings, and they have high tolerance to harsh environmental conditions and proteolytic degradation.

Craniometaphyseal dysplasia (CMD), a rare genetic bone disorder, is characterized by lifelong progressive thickening of craniofacial bones and metaphyseal flaring of long bones. The autosomal dominant form of CMD is caused by mutations in the progressive ankylosis gene ANKH (mouse ortholog Ank), encoding a pyrophosphate (PPi) transporter. We previously reported reduced formation and function of osteoblasts and osteoclasts in a knockin (KI) mouse model for CMD (AnkKI/KI) and in CMD patients. We also showed rapid protein degradation of mutant ANK/ANKH. Mutant ANK protein displays reduced PPi transport, which may alter the inorganic phosphate (Pi) and PPi ratio, an important regulatory mechanism for bone mineralization. Here we investigate whether reducing dietary Pi intake can ameliorate the CMD-like skeletal phenotype by comparing male and female Ank+/+ and AnkKI/KI mice exposed to a low (0.3%) and normal (0.7%) Pi diet for 13 weeks from birth. Serum Pi and calcium (Ca) levels were not significantly changed by diet, whereas PTH and 25-hydroxy vitamin D (25-OHD) were decreased by low Pi diet but only in male Ank+/+ mice. Importantly, the 0.3% Pi diet significantly ameliorated mandibular hyperostosis in both sexes of AnkKI/KI mice. A tendency of decreased femoral trabeculation was observed in male and female Ank+/+ mice as well as in male AnkKI/KI mice fed with the 0.3% Pi diet. In contrast, in female AnkKI/KI mice the 0.3% Pi diet resulted in increased metaphyseal trabeculation. This was also the only group that showed increased bone formation rate. Low Pi diet led to increased osteoclast numbers and increased bone resorption in all mice. We conclude that lowering but not depleting dietary Pi delays the development of craniofacial hyperostosis in CMD mice without severely compromising serum levels of Pi, Ca, PTH, and 25-OHD. These findings may have implications for better clinical care of patients with CMD. © 2020 American Society for Bone and Mineral Research.

In the first part of this publication, the results from an international study evaluating the precision (i.e., repeatability and reproducibility) of N-glycosylation analysis using capillary electrophoresis of APTS-labeled N-glycans were presented. The corresponding results from ultra-high performance liquid chromatography (UHPLC) with fluorescence detection are presented here from 12 participating sites. All participants used the same lot of samples, reagents, and columns to perform the assays. Elution time, peak area and peak area percent values were determined for all peaks ≥0.1% peak area, and statistical analysis was performed following ISO 5725-2 guideline principles. The results demonstrated adequate reproducibility, within any given site as well across all sites, indicating that standard UHPLC-based N-glycan analysis platforms are appropriate for general use.

Based on our empirical evidence, this paper aims to provide a better understanding and analysis of the complexity and diversity of segregated Roma neighbourhoods in the context of Hungarian small towns. The paper explores the differences between the ‘ghetto’ and ‘ethnic neighbourhood’ as typical forms of spatial and social segregation of Roma in Hungary. It also discusses the mutual interplay of spatial and social segregation and social and symbolic boundary‐making within segregated neighbourhoods as well as between culturally and socially diverse Roma groups and non‐Roma living in the given settlements. Special attention is paid to the various social ties between Roma and non‐Roma in these communities. These ties are one of the key elements used to create and maintain ethnic and social hierarchy, further stratifying the unequal access to resources as well as exacerbating exclusion and boundary‐making mechanisms.

Aim: The aim of this work is to test if three single nucleotide polymorphisms (SNPs) implicated in glutamate homeostasis or signalling and cellular survival are associated with birth condition. Methods: This study is drawn from the Avon Longitudinal Study of Parents and Children. 7611 term infants were genotyped and patient outcome data retrieved from routine medical records. Exposure measures were the presence of one or more minor alleles in one of 3 SNPs (rs2284411, rs2498804, rs1835740). The primary outcome was the need for resuscitation at birth. Results: For SNP rs1835740, infants homozygous for the minor allele compared to wild-type were more likely to need resuscitation (9.2% vs. 7.0%, p=0.041) while the odds ratio for resuscitation was associated with each increasing minor allele (OR 1.17 (1.01 to 1.35)). Population attributable risk fraction was 6.5%. There was no evidence that the other two SNPs investigated were associated with birth condition. Conclusions: We have tested three candidate SNPs to measure any association with birth condition. The study revealed that the rs1835740 was associated with the need for resuscitation and Apgar scores, with a substantial population impact. This article is protected by copyright. All rights reserved.

The ursane triterpenoids, asiatic acid 1 and madecassic acid 2, are the major pharmacological constituents of Centella asiatica, commonly known as Gotu Kola, which is used traditionally for the treatment of anxiety and for the improvement of cognition and memory. Using the two-electrode voltage-clamp technique, these triterpenes, and some semisynthetic derivatives, were found to exhibit selective negative modulation of different subtypes of the GABAA receptor expressed in Xenopus laevis oocytes. Despite differing by only one hydroxyl group, asiatic acid 1 was found to be a negative modulator of the GABA-induced current at α1 β2 γ2L, α2 β2 γ2L and α5 β3 γ2L GABAA receptors, while madecassic acid 2 was not. Asiatic acid 1 exhibited the greatest effect at α1 β2 γ2L (IC50 37.05 μm), followed by α5 β3 γ2L (IC50 64.05 μm) then α2 β2 γ2L (IC50 427.2 μm) receptors. Conversion of the carboxylic acid group of asiatic acid 1 to a carboxamide group (2α,3β,23-trihydroxy-urs-12-en-28-amide 5) resulted in enhanced inhibition at both the α1 β2 γ2L (IC50 14.07 μm) and α2 β2 γ2L receptor subtypes (IC50 28.41 μm). The results of this study, and the involvement of α5 -containing GABAA receptors in cognition and memory, suggest that asiatic acid 1 may be a lead compound for the enhancement of cognition and memory.

Fluorescence or Forster resonance energy transfer (FRET) remains one of the most widely used methods for assessing protein clustering and conformation. Although it is a method with solid physical foundations, many applications of FRET fall short of providing quantitative results due to inappropriate calibration and controls. This shortcoming is especially valid for microscopy where currently available tools have limited or no capability at all to display parameter distributions or to perform gating. Since users of multiparameter flow cytometry usually apply these tools, the absence of these features in applications developed for microscopic FRET analysis is a significant limitation. Therefore, we developed a graphical user interface-controlled Matlab application for the evaluation of ratiometric, intensity-based microscopic FRET measurements. The program can calculate all the necessary overspill and spectroscopic correction factors and the FRET efficiency and it displays the results on histograms and dot plots. Gating on plots and mask images can be used to limit the calculation to certain parts of the image. It is an important feature of the program that the calculated parameters can be determined by regression methods, maximum likelihood estimation (MLE) and from summed intensities in addition to pixel-by-pixel evaluation. The confidence interval of calculated parameters can be estimated using parameter simulations if the approximate average number of detected photons is known. The program is not only user-friendly, but it provides rich output, it gives the user freedom to choose from different calculation modes and it gives insight into the reliability and distribution of the calculated parameters. © 2016 International Society for Advancement of Cytometry.

In 2010–2012, new outpatient service locations were established in Hungarian micro-regions, which had lacked such capacities before. We exploit this quasi-experiment to estimate the effect of geographical accessibility on outpatient case numbers using both individual-level and semi-aggregate panel data. We find a 24–27 per cent increase of case numbers as a result of the establishments. Our specialty-by-specialty estimates imply that a 1-min reduction of travel time to the nearest outpatient unit increases case numbers for example by 0.9 per cent in internal care and 3.1 per cent in rheumatology. The size of the new outpatient capacities has a separate effect, raising the possibility of the presence of supplier-induced demand. By combining a fixed-effects logit and a fixed-effects truncated Poisson estimator, we decompose the effects into increases in the probability of ever visiting a doctor on the one hand and an increase of the frequency of visits on the other hand. We find that new visits were dominant in the vast majority of specialties, whereas both margins were important for example in rheumatology. Finally, we demonstrate the usefulness of the fixed-effects truncated Poisson estimator in modelling count data by examining its robustness by simulations. Copyright © 2015 John Wiley & Sons, Ltd.

The Human Immunodeficiency Virus (HIV) envelope glycoprotein (Env) is the primary antigenic feature on the surface of the virus and is of key importance in HIV vaccinology. Vaccine trials with the gp120 subunit of Env are ongoing with the recent RV144 trial showing moderate efficacy. gp120 is densely covered with N-linked glycans that are thought to help evade the host's humoral immune response. To assess how the global glycosylation patterns vary between gp120 constructs, the glycan profiles of several gp120s were examined by capillary electrophoresis with laser induced fluorescence detection and MALDI-MS. The glycosylation profiles were found to be similar for chronic vs. transmitter/founder isolates and only varied moderately between gp120s from different clades. This study revealed that the addition of specific tags, such as the gD tag used in the RV144 trial, had significant effects on the overall glycosylation patterns. Such effects are likely to influence the immunogenicity of various Env immunogens and should be considered for future vaccine strategies, emphasizing the importance of the glycosylation analysis approach described in this paper.

The folding of disulfide proteins is of considerable interest because knowledge of this may influence our present understanding of protein folding. However, sometimes even the disulfide pattern cannot be unequivocally determined by the available experimental techniques. For example, the structures of a few small antifungal proteins (PAF, AFP) have been disclosed recently using NMR spectroscopy but with some ambiguity in the actual disulfide pattern. For this reason, we carried out the chemical synthesis of PAF. Probing different approaches, the oxidative folding of the synthetic linear PAF yielded a folded protein that has identical structure and antifungal activity as the native PAF. In contrast, unfolded linear PAF was inactive, a result that may have implications concerning its redox state in the mode of action.

Multicomponent reactions (MCRs) are extremely popular owing to their facile execution, high atom-efficiency and the high diversity of products. MCRs can be used to access various heterocycles and highly functionalized scaffolds, and thus have been invaluable tools in total synthesis, drug discovery and bioconjugation. Traditional isocyanide-based MCRs utilize an external nucleophile attacking the reactive nitrilium ion, the key intermediate formed in the reaction of the imine and the isocyanide. However, when reactants with multiple nucleophilic groups (bisfunctional reactants) are used in the MCR, the nitrilium intermediate can be trapped by an intramolecular nucleophilic attack to form various heterocycles. The implications of nitrilium trapping along with widely applied conventional isocyanide-based MCRs in drug design are discussed in this review.

Despite their impressive optical properties, lead halide perovskite quantum dots (PQDs) have not realized their potential, especially in bioimaging applications, as they suffer from poor moisture and thermal stability, solvent incompatibility, and significant toxicity. Here, a spray-assisted coil-globule transition method for encapsulating CsPbBr3 (CPB) PQDs into poly(methyl methacrylate) (PMMA) polymer nanospheres is reported. Polyvinylpyrrolidone-capped CPB PQDs are synthesized via the ligand assisted reprecipitation method in dichloromethane. After dissolving PMMA, the above precursor solution is sprayed into petroleum ether under high pressure N2 . High-pressure nebulization restricts the interactions between PMMA polymer chains, resulting in the formation of ≈112 nm nanoscale composite spheres after a coil-globule transition. The CPB@PMMA nanospheres not only possess 73% quantum yields but retain 81% of fluorescence intensity after the exposure to water for over 80 days. Due to their confined size and biocompatible encapsulation, they are readily available for cellular uptake and exhibit no toxicity on live HeLa cells. Furthermore, the PMMA surface allows for functional surface modification, carrying the possibility of targeting specific biological species and processes.

Background and purpose: ABC multidrug transporters (MDR-ABC proteins) cause multiple drug resistance in cancer and may be involved in the decreased anti-cancer efficiency and modified pharmacological properties of novel specifically targeted agents. It has been documented that ABCB1 and ABCG2 interact with several first-generation, small-molecule, tyrosine kinase inhibitors (TKIs), including the Bcr-Abl fusion kinase inhibitor imatinib, used for the treatment of chronic myeloid leukaemia. Here, we have investigated the specific interaction of these transporters with nilotinib, dasatinib and bosutinib, three clinically used, second-generation inhibitors of the Bcr-Abl tyrosine kinase activity.

The eomposition and polymerization activity of a given catalyst system (TiCI4-Et2AlCl) were determined. The solid particles formed by the reaction of the catalyst components were separated at two given times, which were characteristic of the polymerization kinetics. The polymerization activities of the catalyst precipitates were determined in the presence of Et2AlCl and other cocatalysts. It was established that the measured polymerization activities of the catalyst precipitates are higher in the initial system than in the presence of other cocatalysts. The composition of the liquid phase of the catalyst was studied and it was found to contain certain dissolved Ti compounds, the titanium content of which could be determined after hydrolysis as trivalent Ti. The concentration of this titanium compound depends on the solvent used. The polymerization kinetic studies provided evidence that the maximum polymerization rate, i.e., the process of polymerization as a whole, depends on the kind of the solvent used and thus on the quantity of dissolved titanium compounds in the liquid phase.

A cell membrane permeable phosphopeptide corresponding to the SHP-2 binding motif of Grb2-associated binder 1 (Gab1) interferes with the Gab1 adaptor-dependent functions and modulates B cell receptor-triggered intracellular signaling in B cell tumors.

ATP-binding cassette (ABC) transporters are involved in the transport of substrates across biological membranes and are essential for many cellular processes. Of the fifty-six Drosophila ABC transporter genes only white, brown, scarlet, E23 and Atet have been studied in detail. Phylogenetic analyses identify the Drosophila gene dMRP/CG6214 as an orthologue to the human multidrug-resistance associated proteins MRP1, MRP2, MRP3 and MRP6. To study evolutionarily conserved roles of MRPs we have initiated a characterization of dMRP. In situ hybridization and Northern analysis indicate that dMRP is expressed throughout development and appears to be head enriched in adults. Functional studies indicate that DMRP is capable of transporting a known MRP1 substrate and establishes DMRP as a high capacity ATP-dependent, vanadate-sensitive organic anion transporter.

Sexual maturation and sperm release were induced in farmed European eels Anguilla anguilla kept exclusively in fresh water by using two dosages of human chorion gonadotropin (100 International Unit (IU)-Group one and 250 IU/individual per week-Group two). Sperm release took over 13 wk in both groups. The quality of sperm was investigated on the eighth, ninth, and tenth wk. The average cell densities were 0.94 ± 0.4 × 1010 (Group one) and 0.93 to. ± 0.6 × 1010 (Group two) spermatozoa/mL. The estimated motility of eel sperm was 33, 55, and 49% on the eighth, ninth, and tenth wk of treatment, respectively. The estimated average motility of samples selected for cryopreservation was 73 ± 10%, while the post-thaw motility of cryopreserved samples was 36 ± 11%. The extender originally developed for common carp sperm crypreservation together with methanol as cryoprotectant was found suitable for the cryopreservation of European eel sperm.

Many endogenous or xenobiotic lipophilic substances are eliminated from the cells by the sequence of oxidation, conjugation to an anionic group (glutathione, glucuronate or sulfate) and transport across the plasma membrane into the extracellular space. The latter step is mediated by integral membrane glycoproteins belonging to the superfamily of ATP-Binding Cassette (ABC) transporters. A subfamily, referred as ABCC, includes the famous/infamous cystic fibrosis transmembrane regulator (CFTR), the sulfonylurea receptors (SUR 1 and 2), and the multidrug resistance-associated proteins (MRPs). The name of the MRPs refers to their potential role in clinical multidrug resistance, a phenomenon that hinders the effective chemotherapy of tumors. The MRPs that have been functionally characterized so far share the property of ATP-dependent export pumps for conjugates with glutathione (GSH), glucuronate or sulfate. MRP1 and MRP2 are also mediating the cotransport of unconjugated amphiphilic compounds together with free GSH. MRP3 preferentially transports glucuronides but not glutathione S-conjugates or free GSH. MRP1 and MRP2 also contribute to the control of the intracellular glutathione disulfide (GSSG) level. Although these proteins are low affinity GSSG transporters, they can play essential role in response to oxidative stress when the activity of GSSG reductase becomes rate limiting. The human MRP4, MRP5 and MRP6 have only partially been characterized. However, it has been revealed that MRP4 can function as an efflux pump for cyclic nucleotides and nucleoside analogues, used as anti-HIV drugs. MRP5 also transports GSH conjugates, nucleoside analogues, and possibly heavy metal complexes. Transport of glutathione S-conjugates mediated by MRP6, the mutation of which causes pseudoxantoma elasticum, has recently been shown. In summary, numerous members of the multidrug resistance-associated protein family serve as export pumps that prevent the accumulation of anionic conjugates and GSSG in the cytoplasm, and play, therefore, an essential role in detoxification and defense against oxidative stress.

Nearly any aryl or alkyl isocyanide is also effective in transforming (I) to (IV) in low to very good yields.

Finite-element (FE) thermal models have been developed in order to study the temperature distribution in a sliding pair comprising a poly(ether ether ketone) (PEEK) pin and a steel disc in a pin-on-disc configuration. First, a moving heat source model for the disc was created. An alternative distributed heat source model was also produced in order to reduce computing time for the evaluation of the moving heat source model by some orders of magnitude. This latter model gave the same results as the moving heat source model, except for a small region just below the moving heat source. On the basis of the distributed heat source approach, a complete axisymmetric FE model for the disc side (taking the effect of thermal resistance between the assembled components into consideration) and a steady-state quarter model for the pin were developed. Water cooling and air cooling of the steel shaft were also compared. It was found that air cooling allowed a higher temperature in the contact region of the two sliding partners. The experimental results obtained with thermocouples and a thermal camera showed good agreement with the model predictions.

Aquaculture is a multifaceted, dynamic food production sector in Europe. The average annual growth rate of aquaculture production in Western Europe was 5.5% between 1988 and 1998, while in Eastern Europe production declined by 56% during the same period. The main growth in aquaculture production has taken place in the marine environment, particularly in the expanding salmon, Salmo salar L., industry of Northern Europe. Inland aquaculture only contributed 19% of the total aquaculture production in 1998. Trout in Western Europe, and common carp, Cyprinus carpio L., in Eastern Europe are the dominant species in inland aquaculture. Inland fisheries production has been stagnant in Western Europe and has declined considerably in Eastern Europe. The importance of recreational fisheries is increasing all over Europe, although no reliable data are available on angler catches. The major interactions between aquaculture and fisheries are pollution by untreated effluents from farms and impacts on indigenous fish stocks. The conflict is decreasing as more advanced systems are used in inland aquaculture, including water recirculation and effluent treatment. The positive benefit of aquaculture is that the sector supports extensive stocking programmes in commercial and recreational fisheries all over Europe.

Our aim was to set up a protocol in order to provide carrier and prenatal diagnosis to Hungarian haemophilia A (HA) and B (HB) patients and their relatives. For HA, a combination of direct mutation detection and some indirect marker analyses were used: the detection of the inversion mutation and analysis of three polymorphisms, BclI, IVS13 (CA)n and P39(CA)n. In severe cases, direct mutation detection was performed first. In inversion-negative severe cases and in moderate and mild cases, indirect methods were used. For carrier and prenatal diagnosis in HB, four polymorphisms, DdeI, TaqI, XmnI, and HhaI were examined. Our DNA bank contains samples from 50 HA families (34 severe, 15 moderate and one mild) and seven HB families from different parts of the country. In 100% of the HA cases either the gene inversion and/or at least one of the polymorphisms was found to be informative for carrier or prenatal diagnosis. In the HB cases, an informative marker was found in 95% of the cases (19 of 20). We conclude that these strategies are sufficient to make genetic diagnosis available to almost all HA and HB families in the region. This approach is highly informative and cost-effective, so it can be very useful in countries where direct sequencing of genes for factor VIII and IX is not available for routine diagnosis.

Predicting the functional consequence of a given amino acid replacement, that is, the difference between the wild type protein and its nonsynonymous single nucleotide polymorph variants (nsSNPs) is a great challenge. This is especially true in the case of large membrane proteins, like ABC transporters. Kelly et al. has published such an ambitious study in a recent issue of Protein Science focusing on nsSNP in structurally conserved segments within the nucleotide binding domains (NBDs) of ABC transporters supposed to be involved in interdomain communication.1 With the aid of structural rationalization the authors predicted the impact of 40 nsSNPs found in seven clinically important human ABC transporters using a bivalent scoring: disease or neutral. We have performed a deep search of the available literature to find published data on the 40 nsSNPs to delineate them with the predictions. We have incorporated only data from original peer-reviewed publications into our dataset, and our hits were checked in various gene-specific databases too [BioBase and ABCC6 Database]. Based on the available information, we categorized the published data on a given nsSNP as (i) “disease-associated” when the publication clearly demonstrates that the variant segregates with the disease but is absent from at least 200 alleles of unrelated, nonaffected individuals; or (ii) in vitro experimental evidence when the specific variant was heterologously expressed and its (transport) function and/or folding-stability was investigated (Table I). We considered functional differences only if the level of expression of the given nsSNP variant was comparable to the wild type expressed in the same system. Our results, in comparison with the predictions of Kelly et al. are summarized in Table I. The left five columns of the table is taken from the publication of Kelly et al. On the right we list our findings and give the related primary reference. In several cases more than one independent studies were found showing the same result. To keep the list of references short, we indicated only one reference, if more publications were found we added +n (where n is the number of additional independent publications). Table I Predicted and Published Phenotype of nsSNPs in Human ABC Transporters We found the published data on 19 of the 40 ABC nsSNPs. Of these, 16 nsSNPs had been tested in in vitro experiments and in seven examples the given nsSNP resulted in impaired function and/or folding. In one case the published nsSNP generates a new splice site (exon skipping), what affects the mRNA maturation therefore the function or folding of the protein with the given nsSNP can not be concluded (ABCB11, E1186K). One of the nsSNPs was characterized solely by its association to a genetic disease (ABCC6, A1291T associated to pseudoxanthoma elasticum). One additional nsSNP was found as homozygous nsSNP in several healthy individuals therefore it was categorized as neutral, on the basis of this human genetic data (ABCC6, R1268Q). There are two nsSNPs with both published genetic association and in vitro experimental characterization in our data set (ABCB11 V444A associated to intrahepatic cholestasis of pregnancy; ABCG2, Q141K associated to gout). Our conclusions on the nsSNPs are listed in column “Published phenotype” of Table I.; for the sake of simplicity we kept the bivalent terminology of Kelly et al.: disease or neutral. Comparing the predicted phenotype of the Kelly's paper with the phenotype extracted from the published data reveals misprediction in roughly half of the cases (10/19) thus demonstrating the inherent difficulties of rationalizing and predicting the functional impact of snSNPs.

The authors analyzed 1,655 situations from their Genetic Counseling Service over a 15 year period where the reason for counseling was craniospinal anomaly (neural tube defects and/or hydrocephalus) in the family. Excluding the obviously monogenically inherited cases, they investigated pregnancies undertaken after 1,285 isolated and 177 multiple forms of craniospinal abnormalities. The recurrence rate of craniospinal defects was found to be 3.66%, which is about ten times higher than the general population risk, supporting the theory of the multifactorial threshold model in the inheritance of these anomalies. The recurrence risks of neural tube defects and of hydrocephalus were 3.47% and 2.95%, respectively. The authors concluded that recurrence risk is mainly influenced by the pathoanatomic severity of the involved anomaly, the degree of relationship, and the number of affected relatives in the family. There is a positive correlation between the pathoanatomic severity of the anomaly in the proband and the offspring. At least in one-half of the cases the same type of anomaly was observed again in the offspring as in the proband. Attention is drawn to the fact that hydrocephalus (ventriculomegaly) is often manifested only in the second half of gestation. Therefore, performing ultrasound examination is strongly recommended not only at the 18th but at the 24th week of gestation, as well in pregnancies with a positive history of neural tube defects and/or hydrocephalus.

The contact characteristics of spur gears are analysed, taking into consideration real surface micro-geometry, and using numerical algorithms. The contact pressure distribution, contact area, and rigid body rotation of the gears have been calculated for different types of gear surface according to machining: shaping, grinding, or milling. Finally, the sub-surface stress state is evaluated for different pressure distributions, by finite element method.

The influenza virus haemagglutinin has an important role in the infectious cycle of the virus and carries multiple B and T cell epitopes. It is synthesized as a single polypeptide chain but viral infectivity depends on its post-translational enzymatic cleavage. The cleavage site of a trypsin-like enzyme responsible for this modification is found in the most conserved intersubunit region of the molecule. In this study the role of this region in antibody recognition was investigated. Synthetic peptides comprising the intact and cleaved forms of the intersubunit segment were used to examine the specificity of virus- or peptide-induced antibodies. The immune response elicited by viral infection resulted in the appearance of antibodies capable of neutralizing the virus without interfering with its binding to the receptor. A monoclonal antibody (MoAb) of such functional properties was shown to recognize the intact intersubunit region both in the uncleaved haemagglutinin molecule and in a 25-mer synthetic peptide comprising the intact intersubunit region. Specificity and functional studies revealed the conformation-dependent recognition of the C-terminal segment of the haemagglutinin 1 subunit by this MoAb. The binding of the antibody was shown to inhibit the trypsin-mediated cleavage of the haemagglutinin molecule and the membrane fusion event. The enzymatic cleavage of the haemagglutinin was demonstrated to abolish antibody recognition of the infective virus suggesting an escape mechanism mediated by the functional destruction of this highly conserved region. The synthetic peptide corresponding to the intact intersubunit region is characterized by an ordered structure and is able to elicit an antibody response in BALB/c mice while its subfragments are nonimmunogenic. Furthermore, this peptide elicited a protective immune response demonstrated by in vivo experiments.