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Biomedical subjects

H Schneider

Publications and source records attributed to H Schneider.

At least 19 recordsLinked to original sources

Structural analysis of lipooligosaccharide produced by Neisseria gonorrhoeae, strain MS11mk (variant A): a precursor for a gonococcal lipooligosaccharide associated with virulence.

We studied the structure of the lipooligosaccharide (LOS) that is produced by a variant A of strain MS11mk. This variant produces a single LOS that is recognized by monoclonal antibody (MAb) 2-1-L8. In a recent study of the pathogenesis of Neisseria gonorrhoeae in male volunteers, variant A gave rise to other phase variants that produce higher molecular weight LOSs, and these LOS were associated with virulence. Definition of the structure of the variant A LOS is important to understand the biosynthesis of LOS and its expression in vivo. The dephosphorylated oligosaccharide (OS) structure derived from the variant A LOS was analyzed by two-dimensional NMR and methylation analysis. The OS structure was found to be a truncated form of the LOS produced by strain F62 [Yamasaki et al. (1991) Biochemistry 30, 10566-10575]; the variant A OS is a hexamer, a beta-lactosyl residue linked to a tetrasaccharide: Gal beta 1-->4Glc beta 1-->4[GlcNAc alpha 1-->2Hep alpha 1-->3]Hep alpha 1-->KDO. We determined that the variant A LOS is a precursor for the synthesis of higher MW LOS. We also studied expression of the MAb 2-1-L8-defined epitope present on the variant A LOS. Our data indicate that the MAb-defined epitope is not a linear beta-lactosyl residue but its specificity is directed toward the phosphorylated GlcNAc-Hep-Hep residue. Since this MAb binds to gonococci, at least part of the phosphorylated diheptose area is exposed on the gonococcal surface.

Antibodies, Monoclonal

[Acute intermittent porphyria in pregnancy: glucose or hematin therapy?].

The symptoms and management of acute intermittent porphyria (AIP) during pregnancy and delivery are presented on the basis of three case reports. Two patients became asymptomatic after infusion of a concentrated glucose solution (600 ml 50% glucose = 300 g glucose per day), while the third patient needed treatment with hematin (1 mg/kg body weight, 6 infusions at intervals of 12 hours) because of deterioration of psychic state in spite of glucose infusion. Whenever an attack of AIP occurs, concentrated glucose solution should be administrated as a first measure accompanied by an analgesic and possibly a neuroleptic agent after elimination of precipitating factors. However, if symptoms persist for 48-96 hours or the psychic and/or neurological state of the patient further deteriorates, administration of 1-3 mg hematin per kg body weight can be recommended.

Acute Disease

Voltage-gated currents of putative GABAergic amacrine cells in primary cultures and in retinal slice preparations.

To analyze the voltage-dependent ionic conductances of putative GABAergic amacrine cells developing in vitro, whole cell patch clamp recordings were carried out on identified neurons in monolayer cultures from embryonic chick retinae. These recordings were directly compared with those performed on amacrine cells in chick retinal slice preparations. Current responses to depolarizing voltage steps observed in cultured neurons could be separated into at least four different components. A small tetrodotoxin-sensitive sodium inward current was observed in approximately 50% of the cells. The considerably larger outward potassium current consisted of a transient 4-aminopyridine-sensitive component and a sustained component. The latter was reduced in the presence of both tetraethylammonium chloride and Co2+ and thus was probably composed of two conductances. In addition, a Ca(2+)-carried inward current of small amplitude could be identified. Voltage-sensitive currents measured in amacrine cells of retinal slices were very similar. Again, only about half of the cells exhibited sodium currents. Potassium currents contained the above components, but their contributions to the whole cell current seemed to be different. Together with previous findings these results suggest that immature retinal neurons in dissociated cultures undergo a differentiation process similar to that occurring in vivo.

4-Aminopyridine

Lethal Pena-Shokeir 1 syndrome in three male siblings.

A consanguineous family with no living children and three male siblings with Pena-Shokeir 1 syndrome is described. Two children died antepartum and the third shortly after delivery. The importance of early prenatal diagnosis, appropriate counseling and thorough evaluation of the neuromuscular system is discussed.

Abnormalities, Multiple

Creatine phosphokinase isoenzyme activity in umbilical artery, umbilical vein and capillary blood of newborn infants at term.

Serum levels of creatine phosphokinase isoenzyme pattern (CPK MM, MB, BB) were measured in cord blood and capillary blood samples of 20 healthy term infants. Total CPK activity was high but similar in all samples, the main source being derived from the CPK-MM fraction. The CPK-MB fraction was absent in cord blood. Moreover, CPK-BB activity was significantly higher in capillary blood than in cord blood. These results suggest specific organ involvement in the production of CPK fractions.

Creatine Kinase

Effect of thapsigargin on cardiac muscle cells.

The effect of thapsigargin on the activity of various enzymes involved in the Ca(2+)-homeostasis of cardiac muscle and on the contractile activity of isolated cardiomyocytes was investigated. Thapsigargin was found to be a potent and specific inhibitor of the Ca(2+)-pump of striated muscle SR (IC50 in the low nanomolar range). A strong reduction of the Vmax of the Ca(2+)-pump was observed while the Km (Ca2+) was only slightly affected. Reduction of the Vmax was caused by the inability of the ATPase to form the Ca(2+)-dependent acylphosphate intermediate. Thapsigargin did not change the passive permeability characteristics nor the function of the Ca(2+)-release channels of the cisternal compartments of the SR. In addition, no significant effects of thapsigargin on other ATPases, such as the Ca(2+)-ATPase and the Na+/K(+)-ATPase of the plasma membrane as well as the actomyosin ATPase could be detected. The contractile activity of paced adult rat cardiomyocytes was completely abolished by 300 nM thapsigargin. At lower concentrations the drug prolonged considerably the contraction-relaxation cycle, in particular the relaxation phase. The intracellular Ca(2+)-transients elicited by electrical stimulation (as measured by the changes in Fluo-3 fluorescence) decreased in parallel and the time needed to lower free Ca2+ down to the resting level increased. In conclusion, the results indicate that selective inhibition of the Ca(2+)-pump of the SR by thapsigargin accounts for the functional degeneration of myocytes treated with the drug.

Adenosine Triphosphatases

A factor in the increased risk of colorectal cancer due to ingestion of animal fat is inhibition of colon epithelial cell glutathione S-transferase, an enzyme that detoxifies mutagens.

Dietary animal fat increases the risk of colorectal cancer. A factor in the increased risk is hypothesised to result from the inhibition of isoforms of a colonic epithelial cell enzyme that detoxifies genotoxins, glutathione S-transferase, by one of the major secondary bile acids produced in the colon by fat digestion, lithocholic acid. The inhibition allows mutagens to persist in colonic epithelial cells while proliferation is stimulated by secondary bile acids, with a concomitant greater frequency of neoplasia-associated mutations than when proliferation is stimulated in the absence of the mutagens. Elements in the hypothesis include the ability of relatively low concentrations of lithocholic acid to inhibit isoforms of glutathione S-transferase found in colon epithelial cells, entry of lithocholic acid into the epithelial cells, and the correlation of neoplasia-associated colon pathology with high levels of lithocholic acid in fecal water. Higher pH values in the colonic stream are identified as exacerbating the effects of lithocholic acid by increasing its solubility. Lithocholic acid is suggested to be more inhibitory to glutathione S-transferase than the other major colonic secondary bile acid, deoxycholic acid, on the basis of inhibition-structure relationships.

Animals

27Al satellite transition spectroscopy (SATRAS) of polycrystalline aluminum borate 9Al2O3.2B2O3.

27Al NMR spectra of polycrystalline aluminum borate 9Al2O3.2B2O3 have been measured at 104, 130 and 156 MHz. The parameters of the quadrupole interaction and the isotropic chemical shifts have been obtained by fitting the CT/MAS pattern and consideration of the inner satellite transitions m = 3/2<-->1/2 and m = - 1/2<-->- 3/2. The gain in spectral resolution concerned with the observation of the MAS lines of the inner satellites leads to complete separation of the signals of AlO6, AlO5 and AlO4 polyhedra. Also signals of structural groups of one and the same coordination number can be distinguished. Experimental and theoretical lineshape calculations are compared.

Aluminum

Variants of the mucosal mast cell line (RBL-2H3) deficient in a functional membrane glycoprotein.

We have isolated and characterized subpopulations of the rat mucosal mast cell line, RBL-2H3, carrying either high or low density of a glycoprotein, recently established as mast cell function-associated antigen (MAFA, Ortega et al., 1991), on their surface. These populations were investigated in order to better define the involvement of the MAFA in coupling the immunological stimulation of mast cells to mediator release. The MAFA density on the cell surface of the deficient subpopulation was less than or equal to 10-20% that of the parental population and this phenotype was found to be stably maintained for several months. In contrast, the MAFA-enriched cells had maximally twice the number of copies per cell surface than that of the parental population and this phenotype was less stable. Significantly, low copy number of MAFA on the cell's surface was accompanied by a markedly different secretory response, i.e. (i) a considerable decrease in the secretory response to the Fc epsilon RI-mediated stimulus (ii) a marked enhancement of the ionomycin induced secretion. In order to gain insight into the causes for this decrease in cellular response to the Fc epsilon RI-mediated stimulus, we measured the amplitudes of several biochemical processes which are assigned to the stimulus-secretion coupling cascade. The Fc epsilon RI-mediated uptake of 45Ca2+ by the MAFA-deficient cells was considerably lower than that of the parental and MAFA-enriched cells. Similarly, these cell's Fc epsilon RI-induced rise in [Ca2+]i (both the initial transient as well as the sustained elevation), was markedly lower than that of the parental line and the MAFA-enriched cells. Moreover, the low initial transient rise in [Ca2+]i was found to be correlated with the decrease in Fc epsilon RI-mediated IP3 levels. We therefore examined the cell's content of the phosphatidyl-inositides hydrolyzing enzyme, phospholipase C gamma 1. This was found to be similar in the parental line and in its derived subpopulations. However, PLC gamma 1 activation, as measured by the time course of phosphorylation of its tyrosines, showed a marked difference: while PLC gamma 1 tyrosine phosphorylation, in the parental cells, was only transient (detected already 1 min after antigen addition and declined afterwards to basal levels at ca. 10 min), in the MAFA-deficient cells, tyrosine phosphorylated PLC gamma 1 was also observed 1 min after antigen addition, yet showed no decrease with time in its phosphorylation intensity for up to 30 min.(ABSTRACT TRUNCATED AT 400 WORDS)

Aniline Compounds

[Induction of labor with a new intracervically administered prostaglandin E2 gel. Report of initial experience].

We report on our experience with the induction of labour using 0.5 mg of prostaglandin E2 gel (Cerviprost, Organon) intracervically. Out of 43 cases, 27 were primiparae. Indication for induction was: 10 post-term pregnancies, 9 PROM's > 24h, 12 foetal and 11 maternal indications. Mean Bishop score before induction was 3.1 and 9 hours later or at the beginning of regular contractions 6.7. In 9 of the 43 cases induction was unsuccessful. There were 23 spontaneous vaginal deliveries, 6 forceps and 5 Caesarean sections. Uterine hyperstimulation was found in 6 cases, 5 of which occurred during the first 16 inductions, and only one was seen during the following 27. A possible reason may be unrecognised extra-amniotic application of the gel. Intracervical prostaglandin seems to be an appropriate method not only for cervical ripening but also for induction of labour in cases with a low Bishop score. Although side effects are rare, sufficient monitoring and control of mother and foetus are mandatory.

Administration, Intravaginal

Tyrosine phosphorylation of phospholipase C gamma 1 couples the Fc epsilon receptor mediated signal to mast cells secretion.

Mast cells respond to clustering of the type I Fc epsilon receptor (Fc epsilon RI) on their membranes by mediator secretion. Recently, a marked enhancement of tyrosine phosphorylation on several proteins has been observed as a result of antigen-induced Fc epsilon RI aggregation on these cells. We report here that the phosphatidyl inositide specific phospholipase C gamma 1 (PLC gamma 1) is one of the prime proteins that undergoes tyrosine phosphorylation as a result of this stimulus. This was determined by immunoprecipitation of phosphotyrosine containing proteins from detergent lysates of rat mucosal mast cells (rat basophilic leukemia cells, subline 2H3; RBL-2H3) and Western blotting analysis of the separated components. A fast appearance of phosphorylated tyrosine residues on PLC gamma 1 was observed, reaching its maximal intensity at approximately 1-3 min after stimulation and declined afterwards to basal levels. Moreover, the phosphorylation depended on maintaining the aggregated Fc epsilon RI as did other cellular responses (e.g. phosphatidyl inositides hydrolysis and secretion). The time course of both Fc epsilon RI induced phospholipase C gamma 1 activation, as monitored by the formation of inositol phosphates, and of the secretory response of the cells followed that of the PLC gamma 1 phosphorylation. Furthermore, the tyrphostin AG490, a protein tyrosine kinase inhibitor, caused similar inhibition of the Fc epsilon RI-induced PLC gamma 1 phosphorylation, inositol phosphates formation, and mediator secretion. Significantly, no tyrosine phosphorylation of PLC gamma 1 was induced by the Ca2+ ionophore, ionomycin, even at doses that cause optimal secretory response.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Vascular beta-adrenoceptor-mediated responses in hypertension and ageing in rats.

1. In normotensive Wistar (W) and spontaneously hypertensive (SHR) rats between 5 and 20 weeks of age, there was an age-related increase in blood pressure (r2 = 0.770 and r2 = 0.801 respectively). Except for adrenaline (r2 = 0.979) in SHRs, plasma catecholamines and age were unrelated. 2. In ring segments of thoracic aorta from 5, 10, 15 and 20 week old rats, the respective EC80s (-log M) for phenylephrine (PE)-induced contractions were 8.20 +/- 0.37, 7.96 +/- 0.10, 7.15 +/- 0.12 and 7.12 +/- 0.21 in W and 7.73 +/- 0.13, 7.72 +/- 0.16, 7.37 +/- 0.08 and 7.40 +/- 0.03 in SHR tissues (means +/- SEM; n = 5-7). 3. In PE-preconstricted rings, the respective EC50s (-log M) for isoprenaline (IPNA)-induced relaxation were 7.97 +/- 0.11, 7.74 +/- 0.10, 6.96 +/- 0.19 and 6.57 +/- 0.26 in W and 8.03 +/- 0.24, 7.62 +/- 0.08, 6.88 +/- 0.13 and 6.73 +/- 0.14 in SHR tissues (n = 5-7). 4. In PE-preconstricted rings from 5 and 20 week old rats, a single concentration of IPNA (approximating the respective IPNA EC50s) induced relaxation which was sustained over 2 h in W but not SHR tissues. The SHR:W ratios of the net relaxant responses, at 5 and 20 weeks, were 0.6461 and 0.6167 respectively. 5. Thus, W rats exhibit an age-related loss in both vascular alpha- and beta-adrenoceptor responsiveness which appears unrelated to plasma catecholamines. SHRs also exhibit an age-related loss in vasodilator beta-adrenoceptor responsiveness, which may involve adrenaline-induced desensitization, but appear to maintain vasoconstrictor alpha-adrenoceptor responsiveness. It is proposed that an age-related decline in beta-adrenoceptor responsiveness coupled to maintenance of alpha-adrenoceptor responsiveness may lead to chronic blood pressure elevation, as observed in SHRs, while a parallel decline in both alpha- and beta-adrenoceptor responsiveness, as observed in W rats, may preclude hypertension development.

Aging

Rationale for upper limits of electrolyte replacement during exercise.

The addition of carbohydrate and sodium to sport drinks has been recommended to enhance fluid intake and absorption and to delay fatigue. Other electrolytes (E) which are lost through sweating are also commonly added. However, too many E may lead to increased serum E and osmolality levels, which may negatively influence thermoregulation, depress sweating, and cause gastrointestinal distress. On the other hand, drinking large amounts of plain water to compensate sweat loss may induce hyponatremia. Therefore, literature describing sweat E losses was examined in order to estimate average whole-body E loss and to determine an upper limit for replacement of E with sport drinks. Mean E loss was determined from 13 studies, with +/- 1 SD resulting in a hypothetical range for E losses. Correction for net absorption resulted in an upper limit fo electrolyte replacement. It is suggested that the E levels in sport rehydration drinks should not exceed the upper limit of the range given.

Electrolytes

Thermogenesis in obese women: effect of fructose vs. glucose added to a meal.

To assess the effect of a fructose meal on resting energy expenditure (EE), indirect calorimetry was used in 23 women (10 lean and 13 obese) for 30 min before and 6 h after the ingestion of a mixed meal containing 20% protein, 33% fat, and either 75 g glucose or 75 g fructose as carbohydrate source (47%). Expressed as a percentage of the energy content of the meal, the thermogenic response to the fructose meal was significantly greater (10.2 +/- 0.5%) than that of the glucose meal (8.4 +/- 0.4%, P less than 0.01). This difference was still apparent when the lean and obese women were considered separately. The mean respiratory quotient during the 6-h postprandial period was significantly greater (P less than 0.01) for the fructose (0.85 +/- 0.01) than for the glucose meal (0.83 +/- 0.01) in the combined subjects. In addition, cumulative carbohydrate oxidation was significantly greater after the fructose than after the glucose meal (51.1 +/- 2.3 vs. 40.9 +/- 2.0 g/6 h, respectively, P less than 0.01). Only small changes were observed in postprandial plasma levels of glucose and insulin after the fructose meal, but the plasma levels of lactate increased more with fructose than with the glucose meal. These results suggest that there might be some advantages (higher thermogenesis and carbohydrate oxidations) in using fructose as part of the carbohydrate source in diet of people with obesity and/or insulin resistance.

Adolescent