Role of illness representations and coping in patients with atopic dermatitis: a cross-sectional study.

BACKGROUND: Atopic dermatitis (AD) is a skin disease accompanied by psychological burden. It has been shown for other chronic diseases that illness representations and coping strategies are associated with disease-related burden and other outcome variables like time until patients return to work or health care use. OBJECTIVE: The goal of this cross-sectional study was to investigate whether illness representations and coping strategies are correlated with the severity of AD and self-rated physical impairment of the patients. METHODS: A total of 109 AD patients were examined at the beginning of their stay at a rehabilitation centre. They filled in validated questionnaires to measure illness perceptions (IPQ), coping strategies (EBS) and self-rated physical well-being (FEW). In addition, the severity of AD (SCORAD) was determined by a doctor. RESULTS: Linear regression analysis revealed that a considerable amount of the variance in self-rated physical well-being (51%) could be predicted by particular illness perceptions and coping. Subsequent multiple mediation analyses indicated that certain coping strategies (active problem solving and depressive reactions) mediated the effect of illness representations on self-rated physical well-being. In contrast, only 7.4% of the SCORAD could be predicted by the IPQ scale illness identity. CONCLUSION: This study showed that illness representations and coping are highly associated with self-rated physical impairment in AD patients. Therefore, this patient group might profit from cognitive behavioural interventions designed to alter patients' illness perceptions. The hypothesis that a modification in illness perceptions leads to a faster recovery and a more rapid return to work should be tested in future randomized controlled trials.

Quantification of ZP1, ZP2 and ZP3 mRNA of marmoset monkey (Callithrix jacchus) oocytes from periantral and antral follicles.

In mammals, the oocyte and preimplantation embryo are protected by the zona pellucida (ZP) consisting mainly of ZP glycoproteins, which are responsible for sperm binding, induction of the acrosome reaction and zona pellucida hardening to prevent polyspermia. The ZP proteins become increasingly important as possible predictors for in vitro cultured oocytes competence. As little is known about the stage-dependent expression of ZP1, ZP2 and ZP3 in marmoset monkey (Callithrix jacchus) oocytes, mRNA expression was investigated with real-time RT-PCR. Total-RNA was isolated from three different classes of marmoset oocytes; Class 1 oocytes from periantral follicles (<600 µm, n = 10), Class 2 oocytes from small antral follicles (600-1000 µm, n = 10) and Class 3 oocytes from large antral follicles (>1000 µm, n = 9). Compared with Class 1 oocytes mRNA expression of ZP1, ZP2 and ZP3 in Class 2 oocytes was significantly decreased. In Class 3 oocytes, the transcription of ZP1, ZP2 and ZP3 genes showed also a significant decrease compared with Class 1 oocytes. In this study a differently regulated expression of the ZP genes during late folliculogenesis with an obvious downregulation of ZP1, ZP2 and ZP3 could be demonstrated for the first time in the marmoset monkey.

[International Classification of Function, Disability and Health (ICF). Basis for cutaneous rehabilitation management]. / Internationale Klassifikation der Funktionsfähigkeit, Behinderung und Gesundheit (ICF). Grundlage des Handelns und Denkens in der dermatologischen Rehabilitation.

Functioning and health are central issues in medical rehabilitation. With the International Classification of Functioning, Disability and Health (ICF), the World Health Organization implemented in 2001 a system for the description and classification of functioning that is accepted worldwide. Applying the bio-psycho-social perspective, this model based on interdependencies, comprises the components Body Function, Body Structure and Activity and Participation as well as contextual influences of health: the Environmental and Personal factors. The ICF is of prime importance in clinical practice, teaching, and research. It provides a universal language to be used in health care permitting the characterization of the specific functional problems of individuals. The rehabilitation of skin diseases takes advantage on the ICF in rehabilitation management. In this paper we describe the philosophy of the ICF and its implementation in rehabilitation of patients with chronic skin problems on the basis of a case study.

A synthetic decapeptide from a conserved ZP3 protein domain induces the G protein-regulated acrosome reaction in bovine spermatozoa.

In some animal species, the zona pellucida protein 3 (ZP3) plays a central role during fertilization, functioning as a specific receptor for sperm and as an inducer of the acrosome reaction. On the other hand, the zona pellucida protein 2 (ZP2) acts as a secondary receptor, binding to acrosome-reacted sperm. The objective of these studies was to identify ZP2 and ZP3 domains that may be of importance for the induction of the acrosome reaction. For this purpose, we synthesized a number of ZP2 and ZP3 peptides that were either conserved among species or that were species-specific according to their respective primary structures. We identified a defined, conserved ZP3 decapeptide (ZP3-6 peptide) that bound to the surface of the acrosomal region and induced the acrosome reaction in a concentration-dependent manner in capacitated bovine sperm; this effect was significant in the nanomolar range. Pertussis toxin inhibited the ZP3-6 peptide-induced acrosome reaction but had no effect on the progesterone-induced exocytotic event. Our data are in accordance with previous studies showing that progesterone induces acrosomal exocytosis via a different pathway than ZP3 and strengthen the hypothesis that the effect of ZP3-6 peptide upon acrosomal exocytosis is G protein regulated. Despite the commonly accepted idea that glycosylation of ZP proteins is required for successful sperm-oocyte interaction, we found that acrosomal exocytosis can be induced by a synthetic ZP3 peptide that is not glycosylated. The results presented in this study may be useful for the investigation of the molecular mechanisms of sperm-egg interaction in bovine and other species.

Localization and synthesis of zona pellucida proteins in the marmoset monkey (Callithrix jacchus) ovary.

In most species, the zona pellucida (ZP), an extracellular matrix surrounding the mammalian oocyte, is composed of three glycoproteins: ZPA, ZPB and ZPC. Based mainly on results with mice, the site of zona pellucida biosynthesis has been suggested to be exclusively in the oocyte cytoplasm. However, evidence is accumulating that among various species cumulus/granulosa cells may be involved. Because knowledge of ZP biosynthesis in primates is lacking, we used the common marmoset (Callithrix jacchus) to acquire information about the localization and the site of synthesis of ZP proteins in this species. Using antibodies against synthetic ZPA and ZPC peptides, immunoreactivity was found in the marmoset ZP and in surrounding cumulus cells. Interestingly, the amounts of ZPA and ZPC proteins expressed appeared to differ depending on the stage of folliculogenesis. RT-PCR analysis of mRNA from marmoset oocytes and from oocyte-free follicle cells revealed expression of ZPA, ZPB and ZPC in oocytes and in follicle cells of different stages of marmoset monkey folliculogenesis. Our data suggest that the biosynthesis of marmoset ZPA, ZPB and ZPC proteins takes place both in oocytes and in follicle cells of different follicle stages, although the abundance of ZP glycoproteins may differ depending on the individual ZP protein.

Attitudes towards preconception sex selection: a representative survey from Germany.

Within the next parliamentary term, the German government is expected to replace the current Embryo Protection Act with a new Human Reproductive Technology Act. Before introducing new legislation, policy makers may want to survey public attitudes towards novel applications of reproductive technology. In order to assess opinions and concerns about preconception sex selection for non-medical reasons, a social survey has been conducted in Germany. As a representative sample of the German population, 1005 men and women 18 years and older were asked whether or not preconception sex selection should be made available. Of the respondents, 32% held that sex selection should be strictly prohibited, be it for medical or non-medical reasons, and 54% accepted the use of preconception sex selection for medical purposes. Only a minority of 11% approved of the use of preconception sex selection for non-medical reasons. The widespread opposition to a freely available service for non-medical sex selection is based on several claims: 87% of respondents hold that 'children are a gift and deserve to be loved regardless of any characteristics such as beauty, intelligence or sex'; 79% argue that choosing the sex of children is 'playing God'; 76% are opposed because it is seen as 'unnatural'; 49% are afraid that it is 'skewing the natural sex ratio'; and 40% consider it to be 'sexist'.

Preconception sex selection for non-medical reasons:a representative survey from Germany.

BACKGROUND: Preconception sex selection for non-medical reasons raises serious moral, legal and social issues. The main concern is based on the assumption that a freely available service for sex selection will distort the natural sex ratio and lead to a severe gender imbalance. However, for a severe gender imbalance to happen, at least two conditions have to be met. First, there must be a significant preference for children of a particular sex, and second, there must be a considerable demand for preconception sex selection. To ascertain whether or not these two conditions are met, we have conducted a survey in Germany. METHODS: As a representative sample of the German population, 1094 men and women aged 18-45 years were asked about their gender preferences and whether or not they could imagine selecting the sex of their children through flow cytometric separation of X- and Y-bearing sperm followed by intrauterine insemination. RESULTS: 58% of respondents stated that they do not care about the sex of their offspring. 30% wish to have a family with an equal number of boys and girls. 4% would like to have more boys than girls, 3% more girls than boys, 1% only boys and 1% only girls. For first-borns, however, there is still a preference for boys over girls. While 75.6% claimed to have no gender preference, 14.2% would like their first child to be a boy and 10.1% would like their first child to be a girl. Whereas 6% could imagine taking advantage of preconception sex selection, 92% found this to be out of the question. Even in the hypothetical case that a medication for sex selection were ever to become available, 90% stated that they would not want to use it. CONCLUSION: Given that a majority does not seem to care about the sex of their offspring and only a minority seem to be willing to select the sex of their children, a freely available service for preconception sex selection for non-medical reasons is rather unlikely to cause a severe gender imbalance in Germany.

Identification of cytokeratins in bovine sperm outer dense fibre fractions.

Outer dense fibres (ODF) are important substructures of mammalian sperm tails that are involved in the regulation of sperm motility. In this study, we investigated the identity of several sodium dodecyl sulphate (SDS)-insoluble ODF proteins. Bovine ODF were purified by separating sperm heads and tails using ultrasound and Percoll(R) density gradient centrifugation. Sperm flagella were treated with the detergent cetyltrimethylammonium bromide (CTAB). CTAB-insoluble material, which reportedly represents the ODF fraction, was collected, and electron microscopy confirmed a highly purified ODF fraction. We found after solubilization of this fraction with SDS that high amounts of insoluble material were retained after centrifugation. SDS-insoluble material was collected and quantitatively dissolved in 8 M urea. SDS-gel electrophoresis in the presence of urea revealed polypeptides with apparent molecular masses of approximately 25, 43, and 50 kDa. Subsequent immunoblotting with anti-cytokeratin antibodies detected two urea-soluble, SDS-insoluble proteins with apparent molecular masses of 45 and 66 kDa. The 45-kDa protein was identified as cytokeratin 19. An antibody reacting with a palette of cytokeratins (CK 1-18 and CK 20), KL1, was the only antibody that reacted with the 66-kDa polypeptide. We conclude that sperm ODF fractions contain at least one each of type I and type II intermediate filaments. As keratins and intermediate filaments are described as rope-like structures, we suggest that these intermediate filaments play an important structural or tension-bearing role in sperm flagella.

VDAC2 (porin-2) expression pattern and localization in the bovine testis.

In this study, sequencing of voltage-dependent anion channel 2 (VDAC2, porin-2) cDNA from bovine testis is reported. High identity to the murine, rabbit, and human subtypes at both the nucleotide and amino acid levels is demonstrated. mRNA analysis revealed expression of VDAC2 in bovine testis, whereas high levels of VDAC2 proteins were found in late spermatocytes, spermatids, and spermatozoa. In contrast, VDAC1 (porin-1) is exclusively localized in Sertoli cells. The possible role of testicular VDAC2 in providing energy metabolites and in germ cell apoptosis is discussed.

In vitro tests for essential sperm functions using the phyto-oestrogen genistein as a test substance.

Sperm motility, binding of spermatozoa to the zona pellucida and induction of the acrosome reaction are prerequisites for successful oocyte fertilization. Examination of the physiological and nonphysiological effects of particular compounds on sperm functions requires high-quality in vitro test systems. In this short methodological overview, a reliable combined in vitro test system with bovine gametes is described. The purpose of the study was to evaluate whether aliquots of pooled post-thaw spermatozoa are suitable for examination of environmental substances that affect essential sperm functions. The combined test system includes a number of known methods for the assessment of sperm vitality and motion parameters, acrosomal status, inducibility of acrosome reaction and sperm zona pellucida binding. First observations indicate that genistein inhibits the induction of acrosomal exocytosis and binding of spermatozoa to the zona pellucida. Motility parameters and the viability of bovine spermatozoa were not affected by this substance. It is concluded that genistein, a phyto-oestrogen which is abundant in several plants, can be used as a test substance for the evaluation of effects upon essential bovine sperm functions in vitro.

The zona pellucida 'receptors' ZP1, ZP2 and ZP3.

The male component that is necessary for successful reproduction depends on a large variety of biological processes working in concert. The sperm-egg interaction occurs through complementary molecules and is an obligatory process for successful fertilization. However, this complex phenomenon and its molecular mechanisms remain to be fully understood. The oocyte is protected by the zona pellucida, a network of various proteins which encloses the oocyte. Depending on the species, the zona pellucida consists of different glycoproteins that are proposed to function as 'receptors' for spermatozoa. In the mouse, ZP1 is the homodimeric filament crosslinker, held together by intermolecular disulphides. ZP2 is the 'secondary receptor', which is cleaved by egg proteases after egg activation. The mouse ZP3 protein appears to be the 'primary receptor', which is responsible for species-specific binding of spermatozoa to the oocyte and the induction of the acrosome reaction. To localize zona pellucida protein and to evaluate the function of ZP2 and ZP3, polyclonal antisera were raised against synthetic ZP2 or ZP3 peptides which are specific for human or for mouse zona pellucida proteins. It could be demonstrated that anti-synthetic peptide antisera detected their respective zona pellucida proteins in immunoblots, ovary sections and native hemizonae pellucidae. Functional assays with anti-ZP3 synthetic peptide antibodies revealed that the antisera did not inhibit sperm-zona pellucida binding, whereas one of the antisera against synthetic ZP2 peptides significantly inhibited binding of spermatozoa to the zona pellucida.

Species specificity of human and murine anti-ZP3 synthetic peptide antisera and use of the antibodies for localization and identification of ZP3 or ZPC domains of functional significance.

The mammalian zona pellucida has an important function in the fertilization process. The zona pellucida protein 3 (ZP3 or ZPC) is the ligand for primary sperm binding and induces the acrosome reaction. In various species, ZP3 primary structures are highly conserved as revealed by cDNA cloning. The objective of these studies was to localize ZP3 protein using antisera generated against defined synthetic peptides that are specific for mouse or for human ZP3. Immunohistochemistry and transmission electron microscopy were applied to murine and human ovary sections. Immunochemical studies were performed in hemizonae pellucidae from microbisected human oocytes. Using the competitive hemizona assay and various anti-ZP3 antibodies, we further intended to identify human ZP3 epitopes of functional significance. Our results showed that antiserum AS ZP3-9 (mouse specific) detected mouse ZP3 protein in mouse oocytes and in immunoblots, whereas AS ZP3-14 (human specific) detected human ZP3 protein in human ovary sections, native hemizonae pellucidae and in immunoblots. ZP3 material was also detected in cumulus cells by immunohistochemistry. Ultrastructural studies showed an equal distribution of ZP3 throughout the zona pellucida. The human competitive hemizona assay revealed that none of the anti-ZP3 synthetic peptide antisera affected sperm binding suggesting that those epitopes are not involved in primary sperm binding. Anti-porcine ZP3 beta protein antibodies (polyclonal) blocked human sperm-zona pellucida binding. In summary, these anti-ZP3 synthetic peptide antibodies specifically reacted with intact ZP3 protein (murine and human) but did not inhibit human sperm-zona pellucida binding; anti-ZP3 antibodies can therefore be used as biomarkers for ZP3 localization and function.

Corrosion monitoring with speckle correlation.

The changes in the microtopography of a metal surface during a corrosion process are measured by decorrelation of the scattered speckle fields under coherent illumination. For that purpose a quantitative relation between the decorrelation of the scattered light fields and the rate of corrosion is established in a theoretical model, based on the statistics of phase and reflectivity changes of point scatterers at the surface. The speckle fields are recorded by a CCD camera and processed numerically in a computer, yielding the standard deviation of the topography changes with nanometer sensitivity. From the analysis of a series of images taken at equal time intervals during the corrosion process, the degree of interrelation among subsequent topography changes is calculated.

Immunological identification of zona pellucida 2 (ZP2) protein in human oocytes.

The ZP2 protein is a zona pellucida glycoprotein that plays a major role in fertilization. It mediates secondary binding of spermatozoa and is one of the proteins that are involved in zona 'hardening'. ZP2 proteins were identified in various mammalian zonae pellucidae. Their primary structures are highly conserved as revealed by cDNA cloning. Antisera were used against synthetic peptides generated either against a ZP2 amino acid that is homologous in human and mouse ZP2 amino acid sequences (AS ZP2-20) or antibodies against a synthetic human ZP2 peptide (AS ZP2-26). Immunoblots showed that antiserum AS ZP2-20 and AS ZP2-26 strongly recognized human ZP2 protein with an apparent molecular mass of about 72 kDa; both antisera reacted with a minor immunoreactive polypeptide at 96 kDa. In human ovary sections, both antisera revealed immunoreactivity to human zonae pellucidae. Immuno-electron microscopy demonstrated an equal distribution of ZP2 throughout the human zona pellucida. Considerable amounts of immunoreactive material were observed in the ooplasm; some ramification-like extensions of zona pellucida antigen were found close to cells surrounding the oocyte. Our results indicate that antisera against synthetic ZP2 peptides can be used as specific markers for the identification of ZP2 protein in human oocytes.

Evaluation of ZP2 domains of functional importance with antisera against synthetic ZP2 peptides.

The mouse zona pellucida protein ZP2 plays an important role in the process of fertilization by mediating secondary sperm binding to mammalian oocytes. ZP2 primary structures are highly conserved as revealed by cDNA cloning. The aim of the study was to identify ZP2 domains of functional relevance. Antisera were raised against synthetic peptides that are either conserved in the structure of ZP2 from different mammalian species (AS ZP2-20) or present in the human ZP2 but not in the mouse ZP2 amino acid sequence (AS ZP2-26). Antibody binding to zona pellucida proteins was assessed by assaying the antisera with human hemizonae. Using human zonae pellucidae, we demonstrated that anti-ZP2 common antibodies and anti-ZP2 human peptide antibodies react with human zona pellucida antigens. For the first time, ZP2 domains of functional relevance for human sperm-oocyte interaction could be identified applying the competitive hemizona assay. Antiserum AS ZP2-20 significantly inhibited binding of spermatozoa to test hemizonae, whereas treatment of hemizonae with AS ZP2-26 did not influence sperm-oocyte interaction. These results show that antibodies against synthetic ZP2 peptides react with ZP2 protein and that AS ZP2-20 identifies a linear ZP2 epitope that is of possible functional importance for sperm-oocyte interaction.

A new combined in-vitro test model for the identification of substances affecting essential sperm functions.

Binding of mammalian spermatozoa to the zona pellucida and the induction of the acrosome reaction are prerequisites for successful oocyte fertilization. It has been postulated that xenobiotics that are released in the environment as well as exposure to pharmaceutical medications may be associated with reproductive problems in men and wildlife. Examining physiological and non-physiological effects of particular compounds on sperm functions requires high quality in-vitro test systems. We established a reliable combined in-vitro test system with bovine gametes and evaluated if aliquots of pooled post-thaw spermatozoa are suitable for examining essential sperm functions. Using cryopreserved semen, the PSA-FITC/Hoechst 33258 staining procedure was applicable to evaluate the acrosomal status and cell viability. In the bovine hemizona assay, hemizona indices revealed no differences between cryopreserved and fresh semen. Treatment of post-thaw bovine spermatozoa with progesterone (1 microM or bovine follicular fluid (20%) induced the acrosome reaction from 12% (untreated spermatozoa) to 25% (P < 0.001) and to 22% [corrected] (P < 0.01), respectively. Incubation of both compounds (1 microM progesterone and 20% follicular fluid) raised the percentage of acrosome-reacted spermatozoa to 30% (P < 0001). Our results demonstrate that cryopreserved semen can be integrated into an in-vitro screening model for reproductive toxicology testing. Pooled, cryopreserved bovine spermatozoa will thus permit reproducible experiments for clinical and basic science purposes and may also be applicable for the human system.

Spatial phase shifting for pure in-plane displacement and displacement-derivative measurements in electronic speckle pattern interferometry (ESPI).

Hitherto no method, to our knowledge, was known to incorporate spatial phase shifting for the measurement of pure in-plane displacements. We demonstrate that the modified Duffy two-aperture configuration [Opt. Lett. 22, 1958 (1996)], which is sensitive to only the in-plane displacement component and offers increased sensitivity, lends itself to measurement with spatial phase shifting. The configuration can also be used for obtaining displacement derivatives by the introduction of shear with the tilt of a mirror.

Light-in-flight holography for visualization and velocimetry in three-dimensional flows.

Flow visualization and particle image velocimetry of deep volumes are achieved by holographic recordings. Light-in-flight holography is applied to avoid noise from out-of-focus regions during interrogation of the reconstructed image by use of a ruby laser source of small coherence. The scheme permits reconstruction of thin layers in depth without disturbance by the rest of the field. The location of the layers is selected by the position of a reconstructing slit aperture on the hologram. Applications to tracer particles in water and visualization by smoke of an air flow are given.

The zona pellucida "receptors".

Binding of mammalian sperm to the zona pellucida and the induction of the acrosome reaction are prerequisites for successful oocyte fertilization. In the mouse model, the zona pellucida consists of three sulfated glycoproteins, ZP1, ZP2, and ZP3. Zona pellucida proteins are secreted to form a filamentous zona matrix in which ZP2 and ZP3 complex into co-polymers cross-linked by ZP1. ZP3 is the ligand for primary sperm binding and important for the induction of the acrosome reaction. The zona pellucida glycoprotein ZP2 is also crucially involved in the process of fertilization. Previous reports suggest that ZP2 mediates secondary binding of spermatozoa and that cleavage of ZP2 by proteases released through cortical granule reaction causes zona "hardening" and thus prevents polyspermy. Human and mouse ZP2 proteins differ in the primary structure as derived from cDNA clones. We designed an immunological approach to search for ZP2 domains with functional relevance. Antisera were generated against synthetic peptides derived (a) from ZP2 amino acid sequences that are homologous in human and mouse ZP2 amino acid sequences (AS ZP2-20) or (b) from human ZP2 amino acid sequences that differ from the mouse ZP2 sequence (AS ZP2-26). Immunochemical studies with microbisected bovine zonae pellucidae demonstrated that both antisera, AS ZP2-20 and AS ZP2-26, specifically detected ZP2 protein. Using the competition-hemizona-assay, sperm binding to antibody treated bovine hemizonae pellucidae were compared with control hemizonae (given as hemizona index). Antiserum AS ZP2-20 significantly inhibited binding of spermatozoa to test hemizonae (p < 0.0001), whereas treatment of hemizonae with AS ZP2-26 did not influence sperm-egg interaction. Our results show that antibodies against ZP2 peptides react with bovine zonae pellucidae and can be used as markers for ZP2. Furthermore, AS ZP2-20 identifies a ZP2 epitope that is possibly of functional relevance for sperm-egg interaction.