A novel microsatellite polymorphism in the human OB gene: a highly polymorphic marker for linkage analysis

The mouse ob gene and its human homologue OB have recently been cloned. The mutations in the ob gene are known to be associated with extreme obesity. The relationship between the human OB gene and disease, however, is largely unknown due to the lack of suitable markers within or adjacent to the OB gene. To obtain informative markers, we searched for simple tandem repeat polymorphisms in the genomic sequence of the human OB gene and identified a novel tetranucleotide repeat in the 3' flanking region. Fifteen alleles were detected in this marker with a heterozygosity of 0.85 and polymorphism information content of 0.83, indicating a highly informative nature of this marker. Two-point linkage mapping in two Centre Etude Polymorphisme Humaine (CEPH) reference families suggested that this marker is located in the interval between D7S514 and D7S530, the same interval where the OB gene is located (recombination fractions with D7S514 and D7S530 were 0.026 and 0.034, respectively). Although allele frequency distributions of this marker did not differ between 84 control subjects and 69 NIDDM patients, there was a tendency to higher body weight in control subjects with class I/class I genotype than in those without this genotype (68.8 +/- 11.1 vs 60.8 +/- 10.3 kg, p = 0.05). The highly polymorphic nature of this marker and its location in the OB gene makes this marker useful for linkage studies of the OB gene with a number of phenotypes, such as obesity, non-insulin-dependent diabetes mellitus, hypertension and the insulin resistance syndrome.     

The genetic polymorphism of the fourth component of human complement: methodological aspects and a presentation of linkage and association data relevant to its localization in the HLA region

The C4 polymorphism in man has been studied by immunofixation electrophoresis, crossed immunoelectrophoresis, and functional detection after agarose gel electrophoresis. It has so far not been possible to reveal this polymorphism by isoelectric focusing and functional detection of C4 bands. Three common alleles and one less frequently occuring allele have been identified. In a small population sample studied by all the different techniques and verified by family segregation, the following gene frequencies have been found: C4F: 0.46, C4S: 0.32, C4F1: 0.20, and C4M: 0.02. By linkage and association studies in a family material it has been shown that a structural C4 locus is situated in the HLA region of chromosome 6 very close to the HLA-B and Bf loci.     

Multiple sclerosis and the HLA-D region: linkage and association studies

Inheritance patterns of multiple sclerosis (MS) in multiplex families suggest a complex aetiology involving environmental and genetically determined components. The association between the HLA class II DR15, DQ6, Dw2 haplotype and MS has been well documented in patients with ancestral origins in Northern Europe. Conversely, linkage analysis of this region in multiplex families, derived from a population base, has generated negative results. Thus, given the Dw2 specificity association, evidence implicating this locus in disease susceptibility appears contradictory. We have collected and determined the HLA-DR and -DQ haplotypes of 115 sibling pairs with multiple sclerosis, and confirm a significant association with the Dw2-associated haplotype, both in index cases and their affected siblings compared with controls. However, using a sibling pair linkage analysis that restricts haplotype sharing probabilities to defined genetic models, we have not observed linkage of this region to susceptibility in MS. We discuss the basis for association and linkage and conclude that the DR15, DQ6, Dw2 haplotype does represent a susceptibility locus but its contribution to the pathogenesis is small; although it may interact epistatically with other susceptibility genes, this haplotype is not necessary for disease expression.     

Comparative estimation of the life of a material under creep conditions using various approaches

A method for the estimation of the time to failure under creep conditions is proposed; it is based on the approach of a phase transition theory and the similarity of fracture mechanisms. A change in the dominant fracture mechanism is considered as a critical event, and an order parameter is taken to be a reduced time to failure. This parameter reaches its critical values at the stresses corresponding to changes in the fracture mechanisms. The reduced order parameter is related to the reduced stress by a power function, which is used to estimate the time to failure. The accuracy of this method is shown to be higher than those of the well-known time and temperature-time prametric methods of life estimation.     

Variations in the chondroitin sulfate-protein linkage region of aggrecans from bovine nasal and human articular cartilages

Aggrecan-derived chondroitin sulfate (CS) chains, released by beta-elimination, were derivatized with p-aminobenzoic acid or p-aminophenol; radioiodinated; and subjected to graded or complete degradations by chondroitin ABC lyase to generate linkage region fragments of the basic structure DeltaGlyUA-GalNAc-GlcUA-Gal-Gal-Xyl-R (where DeltaGlyUA represents 4, 5-unsaturated glycuronic acid, and R is the adduct), by chondroitin AC lyase to generate the shorter fragment DeltaGlyUA-Gal-Gal-Xyl-R, or by chondroitin C lyase to generate the same fragment when it was linked to a 6-O-sulfated or unsulfated GalNAc at the nonreducing end. Fragments were separated by size using gel chromatography, by charge using ion-exchange chromatography, and by size/charge using electrophoresis and then characterized by stepwise degradations from the nonreducing end by using mercuric acetate to remove all terminal DeltaGlyUA, by bacterial glycuronidase to remove the same residue when linked to unsulfated or 6-O-sulfated GalNAc/Gal, by mammalian 4-sulfatase to remove sulfate from terminal GalNAc 4-O-sulfate, by chondro-4-sulfatase to remove 4-O-sulfate from other GalNAc/Gal residues, and by beta-galactosidase to remove terminal Gal. Results with CS from bovine nasal cartilage aggrecan show that, in nearly all chains, Xyl and probably also the first Gal are unsubstituted, whereas the second Gal is 4-O-sulfated in one CS chain out of five. The first disaccharide repeat is sulfated at C-4 of GalNAc in one chain out of three and unsulfated in the other two. A sulfated first disaccharide is always joined to an unsulfated GlcUA-Gal-Gal sequence. In contrast, CS from human articular cartilage usually has a sulfated first disaccharide repeat. In CS from young human cartilage, sulfate groups are mostly at C-4 of GalNAc in the major part of the chain, but at C-6 in the nonreducing distal portion. In CS from old cartilage, sulfation at C-6 of GalNAc is a major feature from the nonreducing end down to approximately positions 4 and 5 from the linkage region, where GalNAc 4-O-sulfate is common.     

Assessing dissimilarity relations under missing data conditions: Evidence from computer simulations.

The extensive research examining relations between group member dissimilarity and outcome measures has yielded inconsistent results. In the present research, the authors used computer simulations to examine the impact that a methodological feature of such research, participant nonresponse, can have on dissimilarity-outcome relations. Results suggest that using only survey responders to calculate dissimilarity typically results in underestimation of true dissimilarity effects and that these effects can occur even when response rates are high. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Studies on the mechanical resistance of human erythrocytes under in-vitro conditions

Hemolysis and resistance of erythrocytes were studies in-vitro in experiments on human ACD fresh blood and on heparinized fresh blood after oxygen dispersion, resp. mixed oxygen and carbon dioxide dispersion, and subsequent treatment in the hemoresistometer. Blood alkalinity increases under oxygen dispersion, acidity increases under dispersion with mixed gases. The length of the period of dispersion is more important for hemolysis than the change of pH. The hematocrit value influences the behavior of blood resistance. High values imply enhanced hemolysis. There is no significant difference of actual hemolysis values between oxygen and oxygen/carbon dioxide dispersion. A different behavior appears in the calculation of difference value of hemolysis after treatment in resistometer and the respective actual hemolysis.     

Enhancement of gene expression under hypoxic conditions using fragments of the human vascular endothelial growth factor and the erythropoietin genes

PURPOSE: Selective gene expression in response to tumor hypoxia may provide new avenues, not only for radiotherapy and chemotherapy, but also for gene therapy. In this study, we have assessed the extent of hypoxia responsiveness of various DNA constructs by the luciferase assay to help design vectors suitable for cancer therapy. MATERIALS AND METHODS: Reporter plasmids were constructed with fragments of the human vascular endothelial growth factor (VEGF) and the erythropoietin (Epo) genes encompassing the putative hypoxia-responsive elements (HRE) and the pGL3 promoter vector. Test plasmids and the control pRL-CMV plasmid were cotransfected into tumor cells by the calcium phosphate method. After 6 h hypoxic treatment, the reporter assay was performed. RESULTS: The construct pGL3/VEGF containing the 385 bp fragment of the 5' flanking region in human VEGF gene showed significant increases in luciferase activity in response to hypoxia. The hypoxic/aerobic ratios were about 3-4, and 8-12 for murine and human tumor cells, respectively. Despite the very high degree of conservation among the HREs of mammalian VEGF genes, murine cells showed lower responsiveness than human cells. We next tested the construct pGL3/Epo containing the 150 bp fragment of the 3' flanking region in the Epo gene. Luciferase activity of pGL3/Epo was increased with hypoxia only in human cell lines. The insertion of 5 copies of the 35-bp fragments derived from the VEGF HREs and 32 bp of the E1b minimal promoter resulted in maximal enhancement of hypoxia responsiveness. CONCLUSIONS: The constructs with VEGF or Epo fragments containing HRE may be useful for inducing specific gene expression in hypoxic cells. Especially, the application of multiple copies of the HREs and an E1b minimal promoter appears to have the advantage of great improvement in hypoxia responsiveness.     

Phylogenetic relationships of bolitoglossine salamanders: a demonstration of the effects of combining morphological and molecular data sets

We analyzed sequence data for 555 bp of the mitochondrial gene cytochrome b in plethodontid salamanders, taken from 18 ingroup (tribe Bolitoglossini) and 4 outgroup (tribe Plethodontini) taxa. There were 257 variable sites, of which 219 were phylogenetically informative. Sequence differences among taxa exceeded 20%, and there were up to 15% amino acid differences among the sequences. We also analyzed 37 morphological (including karyological) characters, taken from the literature. Data were analyzed separately and then combined using parsimony and likelihood approaches. There is little conflict between the morphological and DNA data, and that which occurs is at nodes that are weakly supported by one or both of the data sets. Treated separately, the morphological and DNA data provide strong support for some nodes but not for others. The combined data act synergistically so that good support is obtained for nearly all of the nodes in the tree. Recent divergences are supported by silent transitions, and older divergences are supported by a combination of morphological, karyological, DNA transversion, and amino acid changes. Eliminating silent changes from the DNA data improves the consistency index and improves some bootstrap and decay index values for several deeper branches in the tree. However, the combined data set with all characters included provides a better supported tree overall. Maximum likelihood and parsimony with all of the data give not only the same topology but also remarkably similar branch lengths. Results of this analysis support the monopoly of the supergenera Hydromantes and Batrachoseps, and of a sister group relationship of Batrachoseps and the supergenus Bolitoglossa (represented in this study one species of the genus Bolitoglossa).     

Ways of using saccharose under conditions of insufficient muscular activity

Under conditions of a limited motor activity an excess of administered saccharose is used not to compensate for the energy expenditures, that are below the control level, as shown by the glycolysis and oxygen uptake data, but to participate in the synthesis processes entering the glucosoaminoglycanes, cholesterol and triglycerides. The increasing amount of lipids is favoured by an activation of the pentose cycle (in the liver, heart, brain), due to which the depot of NADP-H2 in the body increases and the limit of liposynthesis is thus lifted.     

Computer simulation of the dynamics of a human arm and orthosis linkage mechanism

BACKGROUND: A prospective study was conducted to evaluate the clinical usefulness of three-dimensional (3D) surface-shaded maps for routine practice of myocardial perfusion single-photon emission computed tomography (SPECT) by comparison with 2D slices and 2D bull's-eye qualitative analysis. METHODS AND RESULTS: Angiograms were performed on 201 consecutive patients, 155 with coronary artery disease (CAD) and 46 with no significant CAD. One-day 201TI stress/rest-reinjection protocol was performed in 110 patients, and 1-day 99mTc-sestamibi or tetrofosmin stress/rest protocol was performed in 91. The stress protocol was either exercise or dipyridamole (0.56 mg/kg) infusion. Three-dimensional surface maps were obtained by using a threshold for the transaxial data at 50%, 55%, 60%, 65%, and 70% of the maximum pixel value in the first 60 patients. Interpretation of 3D maps was based on the presence of a complete (transmural-looking) perfusion hole within the myocardial wall; doubtful patterns were considered pathologic or normal. Good diagnostic values were found for the 50% to 60% thresholds, but the 60% setting showed the best concordance with multislice and bull's-eye analysis; higher values drastically degraded the specificity. Considering doubtful patterns as normal clarified interpretation and led to a small loss in sensitivity but high gain in specificity. Applied to the whole population, the 3D maps using a 60% threshold provided similar diagnostic value to detect CAD as did conventional and bull's-eye analysis. Moreover, the 3D maps showed a trend toward higher specificity and a proportionally smaller decrease in sensitivity (sensitivity: 92.9%, 90.3%, 89.7%; specificity: 45.6%, 50.0%, 58.7% for tomograms, bull's-eye analysis, and 3D maps, respectively), especially for the detection of left anterior descending and right CAD. Multivessel disease was detected in an identical manner. Three-dimensional maps might improve detection of perfusion defects in the basal regions. However, 3D maps were found to be less sensitive than slices and particularly bull's-eye analysis for the reversibility of stress defects. CONCLUSIONS: Three-dimensional surface display of myocardial perfusion is a valuable independent tool for determining presence, extent, and location of CAD. It can convey useful first-look information to the referring physician, especially through a cine-rotational motion (as done in our practice through use of a floppy disk.     

Polymorphic sites in human mitochondrial DNA control region sequences: population data and maternal inheritance

Short-circuit current (Isc), transepithelial conductance (Gt), electrical capacitance (CT) and the fluctuation in Isc were analyzed in polarized epithelial cells from the distal nephron of Xenopus laevis (A6 cell line). Tissues were incubated with Na+- and Cl--free solutions on the apical surface. Basolateral perfusate was NaCl-Ringer. Agents that increase cellular cAMP evoked increases in Gt, CT, Isc and generated a Lorentzian Isc-noise. The responses could be related to active, electrogenic secretion of Cl-. Arginine-vasotocin and oxytocin caused a typical peak-plateau response pattern. Stimulation with a membrane-permeant nonhydrolyzable cAMP analogue or forskolin showed stable increases in Gt with only moderate peaking of Isc. Phosphodiesterase inhibitors also stimulated Cl- secretion with peaking responses in Gt and Isc. All stimulants elicited a spontaneous Lorentzian noise, originating from the activated apical Cl- channel, with almost identical corner frequency (40-50 Hz). Repetitive challenge with the hormones led to a refractory behavior of all parameters. Activation of the cAMP route could overcome this refractoriness. All agents caused CT, a measure of apical membrane area, to increase in a manner roughly synchronous with Gt. These results suggest that activation of the cAMP-messenger route may, at least partly, involve exocytosis of a vesicular Cl- channel pool. Apical flufenamate depressed Cl- current and conductance and apparently generated blocker-noise. However, blocking kinetics extracted from noise experiments could not be reconciled with those obtained from current inhibition, suggesting the drug does not act as simple open-channel inhibitor.     

A haplotype and linkage disequilibrium analysis of the hereditary hemochromatosis gene region

Monoclonal antibodies were generated against serotonin (5-HT) and the C-terminal portion of the neuronal form of nitric oxide synthase (nNOS), the enzyme producing nitric oxide in neurons. These antibodies were used to compare the distribution of 5-HT- and nNOS-containing neurons in the raphe nuclei of four animal species (rat, mouse, guinea pig, and cat). It was found that the rat was the only species in which the raphe nuclei contain a substantial number of nNOS-immunoreactive (IR) cell bodies. In this species and as observed by other authors, all mesencephalic raphe nuclei contained nNOS-IR cells, the largest group being located in the nucleus raphe dorsalis. The coexistence of nNOS and 5-HT immunoreactivities in these nuclei was visualized by double labeling. In the medulla, the nuclei raphe magnus and obscurus displayed a rather low number of nNOS-IR neurons. In the other species, nNOS-IR cell bodies were found in very low numbers, whatever raphe nucleus was considered. The rostral pole of the nucleus raphe dorsalis and the nuclei raphe magnus and obscurus contained a few nNOS-IR neurons which did not show any coincidence with the 5-HT neurons. In addition, nNOS-IR axons were rare. It is concluded that in the mouse, guinea pig, and cat the involvement of nitric oxide in functions subserved by 5-HT within the raphe nuclei might be minimal.     

Clinical judgment under varied informational conditions: Rorschach, personal data, and best guess.

Requested 105 psychological clinicians to make clinical judgments derived from Rorschach information and personal history data (RPD), personal history data alone (PD), or best guesses (BG) on diagnosis, anxiety, and intelligence. Compared with the BG group, the RPD group was significantly more accurate on anxiety and the PD group on diagnosis (p     

Parasitic systems under the conditions of human pressures (the problems of parasitic contamination)

Antioxidant enzymes are present in sphincter of Oddi nerves and regulate sphincter of Oddi motor function mediated by NO-releasing nerves. Oxygen free radicals (O2-.) produce hydrogen peroxide (H2O2) by the action of superoxide dismutase (SOD). Hydroxyl radical (OH.) has been shown to play an important role as a mediator of H2O2 toxicity. The aims of our study were to determine the effects of H2O2 on sphincter of Oddi motility and if these effects are mediated by OH.. Adult opossums were sacrificed and the sphincter of Oddi removed and placed in a tissue bath containing oxygenated Krebs solution. Force transducers recorded tension in a transverse orientation at two sites along the sphincter of Oddi specimen. H2O2 was added into the tissue bath at concentrations from 0.01 to 0.5%. O2-. radicals were inhibited by the addition of SOD, while OH. was scavenged by the addition of alcohol (ETOH) or dimethyl sulfoxide (DMSO). H2O2 produced a dose-dependent increase in baseline amplitude, frequency, and peak amplitude of contractions. The effect of 0.01% H2O2 on sphincter of Oddi contractile frequency was inhibited by 0.2% ETOH and DMSO, but not by SOD. We conclude that H2O2 has profound effects on sphincter of Oddi motility and that the actions of H2O2 are probably mediated through OH..     

Surgical treatment of portal hypertension with ascites using ileovesicopexy under experimental conditions

Experiments conducted with albino rats have evidenced that under the effect of an atherogenic ration the level of lipids and cholesterol in the blood plasma and hepatic tissue and of sillac acids in the blood plasma increased. In the aortic intima the content of acid mucopolysaccharides was rising, this being attended by a concurrent swelling of the main interstitial substance and of the collagen fibers in the subendothelial layer. An addition of apple pectin or of cellulose to the atherogenic ration deferred the development of the mentioned changes.     

Lymphoepithelial cysts of the pancreas: demonstration of lipid component using CT and MRI

We present two cases of surgically proven lymphoepithelial cyst (LEC) of the pancreas that had a lipid component visualized by CT and MRI. Identification of this component in a pancreatic cystic lesion is a key to favor the diagnosis of LEC or splenic epidermoid cyst over other cystic lesions when the lesion is noted in an elderly patient.