The polymorphisms S503P and Q576R in the interleukin-4 receptor alpha gene are associated with atopy and influence the signal transduction

Interleukin-4 (IL-4) plays a major role in immunoglobulin E (IgE) production. Its signal is conferred to effector cells through binding to the alpha chain of the IL-4 receptor (IL-4Ralpha). We present further evidence for polymorphisms in the IL-4Ralpha gene having an effect on IgE regulation. For two of four common polymorphisms, S503P and Q576R, we found an association with lowered total IgE concentrations (P=0.0008 if occurring together). The polymorphism S503P has not yet been described and is located within the I4R motif of the receptor. In vitro analyses using synthetic peptides of this region showed that the tyrosine kinase Janus kinase 1 (JAK1), as well as IRS-1 and IRS-2 bind to the I4R motif irrespective of the polymorphism or a tyrosine phosphorylation. In vivo immunoassays using T cells of four different groups of individuals (S503/Q576; P503/Q576; S503/R576; P503/R576) revealed that only in case of both polymorphisms the phosphorylation of IRS-1 and IRS-2, but not JAK1 was increased. We found no binding of STAT6 to the I4R synthetic peptides; however, the phosphorylation was reduced in the presence of any of the two polymorphisms, including P503 alone. We discuss possible conformational changes of the receptor leading to the observed effects on the phosphorylation status of IRS-1, IRS-2 and STAT6, in addition to previous findings that Q576R alters STAT6 binding. We conclude that P503 and R576 influence the signal transduction pathways through the IL-4Ralpha, an effect that is magnified by the presence of both polymorphisms. This could explain the observed association effects with lowered total IgE concentrations.     

Cadmiumkonzentrationen im blut von stadt- und inselbewohnern in Schleswig-Holstein

Zusammenfassung Die Cadmiumkonzentrationen im Blut von Kindern and Erwachsenen der Landeshauptstadt Kiel and der Nordseeinsel Pellworm (Schleswig-Holstein, Norddeutschland) wurden gemessen. Die Teilnahme an einem Programm der externen Qualitätskontrolle der Europäischen Gemeinschaft sicherte die Richtigkeit der Analysen. Bei den repräsentativen Stichproben von 35 Kindern aus Kiel and 25 von Pellworm im Alter zwischen 7 und 16 Jahren Bowie 45 Kieler and 53 Pellwormer Erwachsenen lagen alle Meßwerte zwischen 0,1 and 4,0 g Cd/l Blut. Kinder sind in beiden Regionen geringer belastet als Erwachsene. Eine Gefährdung durch das Schwermetall Cadmium konnte bei diesen fur Schleswig-Holstein typischen Bevölkerungs-gruppen nicht festgestellt werden, obwohl die Cadmiumbelastung des Blutes durch Zigarettenrauchen signifikant zunimmt.     

Adhesiveness of human uterine epithelial RL95-2 cells to trophoblast: rho protein regulation

Embryo implantation involves adhesion of trophoblast cells to the epithelial lining of the endometrium. Using an in-vitro model to simulate this initial interaction, we previously reported that attachment of human trophoblast-like JAR spheroids to human uterine epithelial RL95-2 cells provokes a Ca(2+) influx in RL95-2 cells depending on apically localized integrin receptors. Here, we demonstrate that adhesiveness of RL95-2 cells for JAR spheroids, measured by a centrifugal force-based adhesion assay, is dependent on Rho GTPases, most likely RhoA. Cellular expression and distribution of RhoA were studied by fluorescence confocal microscopy, focusing on the localization of RhoA and F-actin within the adhesion sites between JAR and RL95-2 cells. Contact areas contained high amounts of RhoA and F-actin fibres near the plasma membrane. To determine whether Rho GTPases may influence JAR cell binding, we treated RL95-2 cells with Clostridium difficile toxin A, which specifically inactivates Rho GTPases. Toxin A treatment changed the subcellular distribution of endogenous RhoA in RL95-2 cells and altered RhoA and F-actin colocalization. Adhesion of JAR spheroids to RL95-2 cells treated with toxin A was largely suppressed. These data indicate that Rho GTPases, most likely RhoA, play an important role in uterine epithelial RL95-2 cells for trophoblast binding, and suggest that RhoA may be involved in local signalling cascades during early embryo implantation in vivo.     

Acoustic voice analysis by means of the hoarseness diagram.

The hoarseness diagram (Michaelis, Fr?hlich, & Strube, 1998a) has been proposed as a new approach to describe different acoustic properties of voices. To test its performance in the analysis of pathologically disturbed and normal voices five requirements are suggested that should be met by any acoustic voice-analysis protocol to be used in voice research and clinical practice. The hoarseness diagram is then tested with regard to these requirements. Individual voices are found to show a satisfactory localization in the diagram. Aspects of stationarity are discussed in the context of four case studies. The different cases illustrate that changes in the acoustic analysis results are observed if the voice-generation conditions change, whereas results are stationary if phonation conditions do not change. Different pathological voice groups defined on grounds of the specific phonation mechanism are found to map to specific regions of the hoarseness diagram, with differences between group locations being significant. All results can be interpreted without exceptions if the two hoarseness diagram coordinates are taken to reflect the vibrational irregularity of the voice-generation mechanisms on the one side and the degree of closure of the vibrating structures on the other side. The hoarseness diagram and its underlying algorithms are thus shown to constitute a useful approach to acoustic voice analysis in research and clinical practice. The tests themselves demonstrate several application possibilities, including the quantitative monitoring of individual voices.     

An investigation of social and class differences in very-low birth weight outcomes: a continuing public health concern

The objective of this study was to examine the relationship of nonmedical factors, including socioeconomic status, social class, education, race, and social support, to low birth weight. In a case-control study of all resident very-low-birth-weight births between December 1, 1989, and March 31, 1991, mothers completed an extensive survey related to their experience of pregnancy, including prenatal and postnatal care. Cases were defined as very-low-birth-weight (VLBW) infants and were matched to moderately-low-birth-weight and normal-birth-weight infants in race, age, and maternal residence. The hypothesis that social and class factors are more predictive of low birth weight than medical factors alone for women without chronic health problems was supported. Although the degree of the association varies depending on birth weight outcome, race even though addressed through matching--continued to play an important role in birth outcomes. A comparison of logistic model performance with and without the inclusion of social factors indicated the importance these variables play in prediction of birth outcomes. This is one of the few studies undertaken that explicitly investigates impact of patient factors on medical care.     

Exposure to polychlorinated biphenyls and levels of thyroid hormones in children.

As part of an epidemiologic study on exposure to a toxic waste incineration plant we investigated whether blood concentrations of polychlorinated biphenyls (PCBs), lead, and cadmium, as well as concentration of mercury in 24-hr urine samples were associated with thyroid hormone status. As an indication of status, we determined levels of thyroid-stimulating hormone (TSH), free thyroxine (FT(4)), and free triiodothyronine (FT(3)) in children living in households where [less than/equal to] 10 cigarettes were smoked per day. Eight PCB congeners (PCBs 101, 118, 138, 153, 170, 180, 183, and 187) were measured in whole blood samples. Of these, seven congeners (PCB 101 was not detected in any sample) and the sum of all PCB congeners were analyzed as predictors for thyroid hormone status in separate linear regression models adjusted for potential confounders. In addition, the possible effects of cadmium, lead, and mercury on levels of thyroid hormones were examined. Blood concentrations and information on questionnaire data were available for 320 children 7-10 years of age. We found a statistically significant positive association between the mono-ortho congener PCB 118 and TSH as well as statistically significant negative relationships of PCBs 138, 153, 180, 183, and 187 to FT(3). There was no association for the PCB congeners and FT(4). Blood cadmium concentration was associated with increasing TSH and diminishing FT(4). Blood lead and urine concentration of mercury were of no importance to thyroid hormone levels. The results stress the need for future studies on the possible influences of PCB and cadmium exposure on thyroid hormones, particularly in children. These studies should also take neurologic development into account.     

Inhibition of lysosomal degradative functions in RPE cells by a retinoid component of lipofuscin

PURPOSE: To investigate the effect of the lipofuscin component N-retinylidene-N-retinylethanolamine (A2-E) on degradative functions of lysosomes in human retinal pigment epithelial (RPE) cells and to evaluate its mechanism of action. METHODS: A2-E was coupled to low-density lipoprotein (LDL). Human RPE cell cultures were loaded with the A2-E/LDL complex, and controls were run with medium containing LDL alone. To determine whether A2-E accumulated in lysosomes, cells were fractionated in a Percoll gradient, and protein degradation was determined by metabolic labeling and measurement of the release of low-molecular-weight radioactivity. Lysosomal degradation was distinguished from nonlysosomal degradation by inclusion of NH4Cl in the medium. The metabolism of sulfated glycosaminoglycans was studied by radiosulfate incorporation in pulse-chase experiments. Intralysosomal pH was determined using a fluorescent lysosomotropic pH indicator. RESULTS: A2-E accumulated almost exclusively in the lysosomal compartment. Lysosomal protein degradation was reduced in a dose-dependent fashion in A2-E-treated cells. The selectivity of A2-E on lysosomal function was demonstrated by its lack of effect on degradation of extralysosomal protein. Lysosomal glycosaminoglycan catabolism of RPE cells was also strongly inhibited by A2-E. Lysosomal pH was increased by A2-E. CONCLUSIONS: The findings indicate that accumulation of A2-E in RPE cells interferes with lysosomal functions as exemplified by its inhibitory effect on protein and glycosaminoglycan catabolic pathways. The quaternary amine character of the A2-E apparently causes a perturbation of the acidic intralysosomal milieu, resulting in diminished hydrolase action and consequent accumulation of undegraded material. Such mechanism could be operative in retinal diseases associated with excessive lipofuscin accumulation including age-related macular degeneration.