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At least 91 records · Page 5Linked to original sources
PubMed2005

Holistic differential analysis of embryo-induced alterations in the proteome of bovine endometrium in the preattachment period.

During the peri-implantation period, molecular signaling between embryo and endometrium (layer of tissue lining the uterus lumen) is supposed to be crucial for the maintenance of pregnancy. To investigate embryo-induced alterations in the proteome of bovine endometrium in the preattachment period (day 18), we used monozygotic cattle twins (generated by embryo splitting) as a model eliminating genetic variability as a source for proteome differences. One of the twins was pregnant after the transfer of two in vitro produced blastocysts, while the corresponding twin received a sham-transfer and served as a nonpregnant control. The two-dimensional fluorescence difference gel electrophoresis (2-D DIGE) analysis of the endometrium samples of three twin pairs (pregnant/nonpregnant) revealed four proteins with significantly higher abundance (p < 10(-9)) in each sample derived from the pregnant animals: Rho GDP dissociation inhibitor beta; 20 alpha-hydroxysteroid dehydrogenase (20 alpha-HSD); soluble NADP(+)-dependent isocitrate dehydrogenase 1; and acyl-CoA-binding protein. To verify the accuracy of the 2-D DIGE quantification, the abundances of 20 alpha-HSD were quantified by a targeted cleavable isotope-coded affinity tag (ICAT) approach. The mass spectrometry-based ICAT quantification matched perfectly the results obtained by 2-D DIGE quantification, demonstrating the accuracy of our data. These results demonstrate that our model (monozygotic twins) in combination with the appropriate analytical tools is particularly suitable for the detection of the proteins involved in the embryo-maternal interactions.

Albumins↗
PubMed1986

Differential analysis of experimental hypermelanosis induced by UVB, PUVA, and allergic contact dermatitis using a brownish guinea pig model.

In moderately colored guinea-pig skin, UVB, PUVA, and allergic contact dermatitis were shown to induce hyperpigmentation that resembled the pigmentary changes observed in mongoloid human skin. Using this model, we examined the effects of chemical agents, including tyrosinase inhibitors and sunscreen agents, on the color changes induced by UV irradiation. The daily exposure of brownish guinea-pig skin to UVB irradiation at a variety of energies for 3 successive days induced clearly visible black pigmentation on the irradiated rectangular areas of the flank within a few days of irradiation, the maximum being reached about 1 week after irradiation, i.e., similar to the changes that occur in pigmented human skin. Split epidermal sheets prepared from untreated pigmented guinea pigs exhibited 200-400 melanocytes/mm2; 1 week after UV irradiation, the applied areas show an increased number of strongly dopa-positive melanocytes with stout dendrites (800-1,000 cells/mm2). UVA irradiation following an intraperitoneal injection of 8-methoxypsoralen (8-MOP) also produced black pigmentation 1 week after irradiation, and this was paralleled by a marked increase in the number of dopa-positive melanocytes in dopa-reacted split epidermal sheets. Allergic contact dermatitis produced by the application of 1-phenylazo-2-naphthol induced hyperpigmentation after an interval of about 14 days in 10 of the 21 allergy-acquiring animals examined. This induced pigmentation was accompanied by an increase in the number of dopa-positive melanocytes as compared to the number seen in controls. In contrast, allergic contact dermatitis produced by the application of dinitrochlorobenzene failed to induce such a high ratio of postpigmentation, with only 3 of the 21 allergy-acquiring animals showing hyperpigmentation and 5 showing depigmentation; in the latter, there was a slight decrease in the number of dopa-positive melanocytes. To study the preventive effect of tyrosine inhibitors on UVB-induced pigmentation, daily topical applications of these compounds were performed after three daily UVB irradiations. Treatment with 10% hydroquinone for 10 days interrupted UVB-induced pigmentation and resulted in a marked reduction in the number of epidermal melanocytes as compared to the number found in UVB-irradiated, untreated control skin.

Animals↗
PubMed2003

Differential analysis of phosphorylated proteins in resting and thrombin-stimulated human platelets.

Blood platelets are important components of haemostasis. After their activation they cause healing of wounds by forming plugs and initiate repair processes. One important event in regulating this activation is the phosphorylation/dephosphorylation of multiple proteins on various tyrosine, serine and threonine residues. To understand the exact molecular mechanisms in platelet activation it is essential to identify proteins involved in the signalling pathways and to localise and characterise their phosphorylation sites. After treatment with (32)P and separation by 2D-PAGE using different pI ranges, phosphorylated platelet proteins were detected by autoradiography. Phosphotyrosine-containing proteins were assigned by immunoblotting with an anti-phosphotyrosine antibody. Another approach for the identification of phosphorylated proteins was immunoprecipitation of tyrosine-phosphorylated proteins using an anti-phosphotyrosine antibody. Protein spots/bands of interest were excised from the gel, digested with trypsin and analysed by MALDI-TOF-MS and nano-LC-ESI-MS/MS, respectively. Several phosphorylated proteins could be identified and the localisation of some in vivo phosphorylation sites was possible.

Amino Acid Sequence↗
PubMed1988

Estimation of binding parameters by kinetic data analysis: differentiation between one and two binding sites.

A method that enables the discrimination between binding models and the estimation of binding parameters, based solely on kinetic data, is described. Experimental data from association and dissociation experiments were fitted simultaneously to models with mono- or biphasic kinetics with the aid of a non-linear maximum likelihood computer program. Discrimination between two models can be performed statistically. The protocol was used to study the binding of the antitussive [3H]noscapine to guinea pig brain homogenate. Two binding processes could be discriminated by their kinetics, despite the fact that [3H]noscapine apparently binds to one homogeneous population of binding sites in equilibrium binding experiments. This method might find general application when two populations of binding sites are suspected from kinetic data, but when selective ligands are lacking. Since parameter estimates are obtained independent of equilibrium binding data, our approach could also serve as an independent control of such experiments, with respect to both Kd and Bmax.

Animals↗
PubMed1988

Differentiated analysis of the secondary structure of hydrophilic and hydrophobic regions in alpha- and beta-subunits of Na+,K+-ATPase by Raman spectroscopy.

Raman spectra of active Na+,K+-ATPase from pig kidney and membrane-bound products of its two-stage trypsinolysis, including alpha-subunit hydrophobic regions as well as the intact beta-subunit and hydrophobic regions of alpha- and beta-subunits, were measured to calculate the secondary structure of hydrophilic and hydrophobic regions of the enzyme. Consequent comparison demonstrated unambiguously that (i) membrane-bound hydrophobic parts of polypeptide chains of Na+,K+-ATPase subunits are in the alpha-helical conformation; (ii) essential contents of the alpha-helix as well as beta-sheet are estimated to form the hydrophilic (mainly cytoplasmic) domain of the Na+,K+-ATPase alpha-subunit; (iii) the exoplasmic hydrophilic domain of the beta-subunit is shown to include several antiparallel beta-pleated sheets and a small amount of the alpha-helix and unordered conformations. The model of the secondary structure organization of hydrophilic domains as well as 8 hydrophobic transmembrane segments of the enzyme molecule was proposed on the basis of experimental results and predictional calculations.

Animals↗
PubMed1978

Quantitative nuclear image analysis: differentiation between normal, hyperplastic, and malignant appearing uterine glands in a paraffin section. III. The use of texture features for differentiation.

Markov chain statistic based texture features were used to discriminate nuclei from three regions in a paraffin section of human endometrium. The data of the digitized nuclei from which the texture features were calculated were the same as described previously (Baak and Diegenbach, 1977; Diegenbach and Baak, 1977). Several texture features proved to give a good separation between two of the three groups of nuclei. Combining two or more features can make a separation of all three groups possible. Most of the features are correlated, indicating that fewer than the 20 features used are needed for discrimination. The results are much better than with the features described in the previous articles.

Cell Nucleus↗
PubMed2006

Differential analysis of D-beta-Asp-containing proteins found in normal and infrared irradiated rabbit lens.

Although proteins are generally composed of l-alpha-amino acids, d-beta-aspartic acid (Asp)-containing proteins have been reported in various elderly tissues. Our previous study detected several d-beta-Asp-containing proteins in a rabbit lens derived from epithelial cell line by Western blot analysis of a 2D-gel using a polyclonal antibody that is highly specific for d-beta-Asp-containing proteins. The identity of each spot was subsequently determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and the Ms-Fit online database searching algorithm. In this study, we discovered novel d-beta-Asp-containing proteins from rabbit lens. The results indicate that beta-crystallin A3, beta-crystallin A4, beta-crystallin B1, beta-crystallin B2, beta-crystallin B3, gamma-crystallin C, gamma-crystallin D, and lambda-crystallin in rabbit lens contain d-beta-Asp residues. Furthermore, the occurrence of d-beta-Asp residues increases with infrared ray (IR) irradiation. Additionally, some d-beta-Asp-containing proteins only appear after IR irradiation. One such protein is the alpha-enolase, which shows homology to tau-crystallin.

Animals↗
PubMed2004

Hair analysis differentiates chronic from acute carbamazepine intoxication.

This is a report of a 12-year-old epileptic child undergoing chronic treatment with carbamazepine who was found comatose. He was considered to have acute severe drug toxicity. Measurement of carbamazepine concentration in the patient's hair segments together with the carbamazepine blood levels were both important in determining the chronic nature of the patient's intoxication.

Acute Disease↗
PubMed2006

Differential analysis of donor characteristics for pancreas and islet transplantation.

Suitable selection of donors is key to the success of human islet isolation and transplantation. Although several important donor-related factors have been identified previously, they needed to be confirmed in our setting. The aims of this study were: (1) to compare the characteristics of islet donors with those of pancreas donors (national transplant registry). (2) to compare the characteristics of islet donors resulting in a successful isolation in our facility with the characteristics of pancreas donors, and (3) to compare the characteristics of islet donors at this facility, whether or not isolation was successful, with donors elsewhere whose islets were transplanted and included in the Collaborative Islet Transplant Registry. The 35 islet isolations completed at our facility were analyzed for various characteristics. Significant differences were seen in donor age body mass index (BMI), and body weight between our islet donors and our pancreas donors (P < .001). These differences were maintained in the subgroup analysis corresponding to donors of successful isolations compared to pancreas donors (P < .01). Most successful isolations in our islet isolation facility were associated with donors of BMI >25. The percentage of successful isolation (>300,000 IEq) was higher among donors with a body weight >90 kg. We concluded that there was little overlap between the donor profiles for pancreas transplantation and for islet transplantation. More specific selection criteria relative to both BMI and body weight for islet donors may result in greater success of pancreas islet isolation and transplantation.

Adult↗