Strain splitting of nitrogen acceptor levels in ZnSe.
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Biomedical subjects
Publications and source records attributed to H Mayer.
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The O-specific polysaccharide of Proteus mirabilis O28 was found to contain D-galactose, D-galacturonic acid (GalA), 2-acetamido-2-deoxy-D-glucose, L-serine, L-lysine, and O-acetyl groups in molar ratios 1:2:1:1:1:1, the amino acids being linked via their alpha-amino group to the carboxyl group of GalA. The polysaccharide was studied using 1H- and 13C-NMR spectroscopy, including selective spin-decoupling, one-dimensional total correlation spectroscopy, two-dimensional homonuclear correlation spectroscopy (COSY), heteronuclear 13C,1H COSY, one-dimensional NOE, and two-dimensional rotating-frame NOE spectroscopy and partial acid hydrolysis followed by borohydride reduction, methylation, and GLC/MS analysis of the derived glycosyl alditols. The following structure of the repeating unit was established: [formula: see text] Epitope specificity of the P. mirabilis O28 polysaccharide was analysed using a homologous rabbit polyclonal antiserum in quantitative precipitation, passive immunohemolysis, and inhibition of passive immunohemolysis. Study with related synthetic glycopolymers (2-acrylamidoethyl glycosides of amides of alpha-D-GalA with amino acids copolymerised with acrylamide) showed the importance of D-GalA(L-Lys) for manifesting serological specificity of the O-antigen. Serological cross-reactions between P. mirabilis O28, S1959, and R14/S1959 (a transient-like form) are discussed.
Human parathyroid hormone (hPTH) has been bacterially expressed in bioreactors as cro-beta-galactosidase-hPTH fusion protein. We have developed a large-scale purification scheme that exploits the pH-dependent differential solubility of hPTH and a two-step chromatographic procedure. We demonstrate that in a number of assay systems, the recombinant material obtained by this procedure is biologically active.
The O-specific polysaccharide, liberated by mild acid hydrolysis of the lipopolysaccharide (LPS-P) from Thiobacillus sp. IFO14570 as isolated from the phenol phase after phenol-water extraction, is shown to have a linear trisaccharide repeating-unit containing three different diamino sugars, namely 2,4-diacetamido-2,4-dideoxyglucuronic acid, 2-acetamidino-4-acetamido-2,4,6-trideoxyglucopyranose, and 2,3-diacetamido-2,3-dideoxyglucuronic acid in the molar ratio of 1:1:1. On the basis of 1H and 13C spectroscopy, including 2D COSY and coherent transfer (RCT)COSY, 1D NOE in the difference mode, and 2D rotating frame NOE(ROESY) the sequence, the type of substitution, and the position of the acetamidino group could be determined. These experiments allowed formulation of the following structure for the O-specific polysaccharide, although the depicted D configuration of the three sugar residues is not yet proven: [structure: see text]
Phenol-water extraction of Rhizobium loti NZP2213 cells allowed a simultaneous isolation of two structurally different lipopolysaccharides from the aqueous (LPS-W) and phenol (LPS-P) phase that differed in their sodium deoxycholate-PAGE pattern and composition. LPS-W showed a profile indicating an R-type LPS; LPS-P had a cluster of poorly resolved bands in the high-molecular-weight region. LPS-P contained large amounts of 6-deoxy-L-talose (6dTal), and a small amount of 2-O-methyl-6-deoxy-talose (molar ratio approximately 30:1), both of which were completely absent in LPS-W. Methylation analysis gave only one major product, 2,4-di-O-methyl-6dTal, indicating that the O-chain is composed of a homopolymer of 1,3-linked 6dTal, having the methylated 6dTal (2-O-Me-6dTal) probably localized at the non-reducing end of the O-chain. This homopolymeric O-chain was additionally O-acetylated, as evidenced by GC-MS and by 13C NMR analysis. The lipid A moieties of both LPS-W and LPS-P showed almost identical composition, with six different 3-OH fatty acids and with two, so far not described, long-chain 4-oxo-fatty acids, all being amide-linked, and with 27-OH-28:0 as the main ester-linked fatty acid. Lipid A was of the lipid ADAG-type, i.e., having a (phosphorylated) 2,3-diamino-2,3-dideoxy-D-glucose-containing lipid A backbone. Lipid ADAG is widespread among species of the alpha-2 group of Proteobacteria, but has so far not been encountered in any other rhizobial or agrobacterial species.
The structural elucidation of lipid A of the cell wall lipopolysaccharide (LPS) of Rhodospirillum salinarum 40 by chemical methods and laser desorption mass spectrometry revealed the presence of a mixed lipid A composed of three different 1,4'bisphosphorylated beta (1 --> 6)-linked backbone hexosaminyl-hexosamine disaccharides, i.e. those composed of GlcN --> GlcN, 2,3-diamino-2,3-dideoxy-D-Glc-(DAG --> DAG, and DAG --> GlcN. Lipid A of R. salinarum contained preferentially 3-OH-18:0 and 3-OH-14:0 as amide-linked and cis delta 11-18:1 and c19:0 as ester-linked fatty acids. The mass spectra of the liberated acyl-oxyacyl residues proved the concomitant presence of 3-O-(cis delta 11-18:1)-18:0 and 3-O-(c19:0)-14:0 as the predominating diesters in this mixed lipid A. The glycosidically linked and the ester-linked phosphate groups of the backbone disaccharide were neither substituted by ethanolamine, phosphorylethanolamine, nor by 4-amino-4-deoxy-L-arabinose, in contrast to most of the enterobacterial lipid As. In the core oligosaccharide fraction, a HexA (1 --> 4)HexA(1 --> 5)Kdo-trisaccharide was identified by methylation analysis. The terminal HexA (hexuronic acid) is possibly 4-OMe-GalA, a component described here as an LPS constituent for the first time. LPS of R. salinarum showed a lethality in C57BL/10 ScSN (LPS-responder)-mice) of an order of 10(-1)-10(-2) of that reported for Salmonella abortus equi LPS, and it was also capable of inducing TNF alpha and IL6 in macrophages of C57BL/10ScSN mice.
In the search of PTH agonists, PTH fragments were examined for both cAMP-dependent and -independent actions on bone-derived assay systems. In a periosteal cell culture system from the embryonic chick, hPTH (18-48) was capable of inducing cell proliferation in a similar manner to hPTH (1-34) and hPTH (28-48) at picomolar concentrations, whereas hPTH (53-84) was inactive. Near physiological concentrations were needed for the proliferative action of PTH which seems to argue against a role for cAMP as a main mediator for growth response. In the tibiae organ culture system of embryonic chick cAMP stimulation and calcium mobilization requires micromolar concentrations. hPTH (18-48) produced, at a concentration of 10(-6) mol/L, a 14-fold increase in cAMP synthesis and an 11-fold increase in calcium mobilization and was 100-fold less active than hPTH (1-34). hPTH (28-48) and hPTH (53-84) had no significant effect. These results demonstrate that hPTH (18-48) retains the ability to induce cell proliferation and exhibits partial agonist activity in the cAMP/PKA signal transduction pathway.
OBJECTIVE: In many retrospective and prospective observational studies, laparoscopic cholecystectomy (LC) compares favorably with conventional cholecystectomy (CC), with respect to length of hospital stay, postoperative pain, and pulmonary function, indicating a diminished operative trauma. Comparison of laboratory findings (stress hormones, blood glucose, interleukins) are a possibility to objectify stress and tissue trauma of laparoscopic and conventional cholecystectomy. SUMMARY BACKGROUND DATA: Major body injury, surgical or accidental, evokes reproducible hormonal and immunologic responses. The magnitude of many of these changes essentially is proportional to the extent of the injury. METHODS: In a prospective study, biochemical stress parameters were measured in the blood of patients undergoing elective cholecystectomy because of symptomatic cholecystolithiasis. Patients with acute cholecystitis, pancreatitis, choledocholithiasis, or malignant disease were excluded. Values from 40 patients after LC and from 18 patients after CC were compared. Both groups had similar patient characteristics, baseline values, and perioperative care, except for deeper anesthesia during CC. RESULTS: On postoperative day 1, epinephrine (p = 0,05), norepinephrine (p = 0.02), and glucose (p = 0.02) responses were higher after CC. Two days postoperatively, norepinephrine remained higher after CC (p < 0.01). Interleukin-1 beta responses were higher during (p < 0.01) and 6 hours after CC (p = 0.03). Interleukin-6 responses were higher 6 hours (p = 0.03), 1 day (p = 0.02), and 2 days (p < 0.01) after CC. CONCLUSIONS: The results show significant lower values of intraoperatively and postoperatively measured epinephrine, norepinephrine, interleukin-1 beta, and interleukin-6 in patients with laparoscopic cholecystectomy, indicating a minor stress response and tissue trauma in this group of patients. The results correspond to the favorable results of most other trials evaluating clinical aspects of laparoscopic cholecystectomy.
The aim of this study was to investigate the role of body fat distribution on steroid hormone serum concentrations in obese adolescent girls before and after weight reduction. Ninety-two girls (age, 15.1 +/- 0.7 yr) with a mean body mass index of 31.2 +/- 4.6 kg/m2 participated in this 6-week intervention study. Initially, girls with abdominal obesity (waist to hip ratio, > 0.86; n = 30) had higher levels of total and free testosterone and lower levels of sex hormone-binding globulin as well as lower morning levels of total and free cortisol than girls with gluteal-femoral obesity (waist to hip ratio, < 0.80; n = 31) independent of their body mass index. After a mean weight loss of 8.3 +/- 2.6 kg by a standardized weight loss program, significant reductions were observed in estradiol, total and free testosterone, dehydroepiandrosterone sulfate, and the ratio of LH to FSH, whereas sex hormone-binding globulin and free cortisol levels increased significantly. Decreases in total and free testosterone and increases in total and free cortisol were significantly greater in the girls with abdominal obesity than in the girls with gluteal-femoral obesity. Our results suggest that obese girls with an abdominal pattern of fat distribution exhibit more pronounced steroid hormone aberrations, in particular a high androgenic activity, than girls with a gluteal-femoral pattern of fat distribution. The reduction of excess body weight by a conventional treatment regimen is associated with a remarkable improvement of steroid hormone abnormalities in this particular subtype of obese adolescent girls.
Lipoprotein(a) [(Lp(a)], a low-density lipoprotein (LDL)-like particle, contains in addition to LDL a specific protein component, apolipoprotein(a) [apo(a)]. Conventionally, Lp(a) has been measured by immunological methods that distinguish between Lp(a) and LDL by dealing with apo(a) as an antigen. We describe a new method to determine Lp(a) on the basis of its cholesterol content. Very-low-density lipoproteins were removed from serum by preparative ultracentrifugation at a density of 1.006 kg/L. The infranate was subjected to agarose gel electrophoresis to separate Lp(a) and LDL. Lp(a) cholesterol was then determined by direct enzymatic staining for cholesterol. On electrophoresis of the > 1.006 kg/L (bottom) fraction, Lp(a) migrates to the pre-beta position, regardless of the genetic apo(a) isoform. The interassay CVs of Lp(a) cholesterol determinations ranged from 6.9% to 11.5%, and the results correlated well with the Lp(a) concentrations measured by immunonephelometry (r = 0.937). There was an inverse relation between the molecular mass of the genetically determined apo(a) isoforms and Lp(a) cholesterol concentrations. Patients with angiographically proven coronary artery disease (CAD) had significantly more Lp(a) cholesterol than healthy controls did. The ratio of Lp(a) cholesterol to immunologically determined Lp(a) tended to be lower in CAD patients, suggesting that Lp(a) particles contained less cholesterol than apo(a). In addition, the new method allows determination of LDL cholesterol without contamination by Lp(a).
Polysaccharides, isolated from the lichen Cetraria islandica, have antimicrobial effectiveness. For pharmaceutical applications the two glucan components lichenan and isolichenan as well as the galactomannan component are of actual interest. Especially the a-glucan isolichenan ist used as an active ingredient in cough lozenges. The conditions for the extraction of the raw material, mainly pH and temperature, have a strong influence on the yield of lichenan, isolichenan, and galactomannan, and also on the amount of tannins in the extract. Target products and also by-products give higher extraction yields with increasing extraction temperatures. Hot water extraction with subsequent fractionation of the extracted polysaccharides by multiple freezing/thawing steps and water removal applying ethanol and ether permitted the isolation of the target polysaccharides in preparative quantities. Tannins were removed by reversed phase chromatography. IR and NMR spectroscopy were used for structural characterization of lichenan and isolichenan. After optimization of the hot water extraction process no significant lower extraction and fractionation yields have been obtained compared to the established tricky DMSO extraction procedure.
Lipopolysaccharide (LPS) was isolated from Thiobacillus ferrooxidans IFO 14262 by the hot phenol-water extraction procedure. The O-specific polysaccharide, liberated from LPS by mild acetic acid hydrolysis, had a branched pentasaccharide repeating-unit composed of D-glucose, L-rhamnose, D-rhamnose, and 3-O-methyl-L-rhamnose in approximate molar ratios of 2:1:1:1. On the basis of methylation analysis, 1H and 13C NMR spectroscopy, including 2D shift-correlated (COSY) and 1D NOE spectroscopy, the structure for the repeating unit of the O-specific polysaccharide was established, and the assumed biological repeating unit indicated.
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STUDY DESIGN: A group of 160 incumbent male railroad workers volunteered for a study of lumbar spine physical capacity. METHODS: They were initially subjected to inclinometric measurements of sagittal and coronal regional mobility (T12-S1). They also were tested on isokinetic trunk strength measurement devices for sagittal (bending) and axial (twisting) trunk strength at multiple speeds. RESULTS: Results showed that they demonstrated mild deficits of lumbar sagittal extension mobility, trunk extensor strength, and sagittal/axial high speed (150-180 degrees/sec) trunk strength when compared with population averages from a previously derived normative database of a nonhomogeneous male population (matched for age and body weight). CONCLUSIONS: It was hypothesized that instructions about "correct lifting techniques," uniformly provided to these workers, may have resulted in an unintended decrement in trunk mobility and strength.
STUDY DESIGN: Quantified cervical physical examination data are provided for a group of 160 incumbent railroad workers from four laboring crafts. OBJECTIVES: The subjects were tested for cervical mobility and strength, according to standardized protocols as part of a comprehensive physical examination and functional testing procedure to establish normative data associated with the axial musculoskeletal system. RESULTS: Tri-planar cervical inclinometric mobility norms are provided, as are sagittal and coronal isometric cervical strength data.
STUDY DESIGN: A group of 160 incumbent male railroad workers was administered a battery of isokinetic and isoinertial lumbar/cervical lifting tests that served as a paradigm for whole-person functional testing of manual handling tasks. RESULTS: Results demonstrated that the workers' performance was near normal or somewhat above population averages according to previously derived heterogeneous normative samples. However, there were some differences among the four laboring crafts that made up the present incumbent worker sample. CONCLUSIONS: The implications of these differences are discussed.
The inner-core region of the lipopolysaccharide of an UDPGalNAc-4-epimerase-deficient mutant of Yersinia enterocolitica 0:3, designated as Ye75R, was investigated using methylation analysis, 1D-13C-NMR and 2D-13C-NMR and 1H-NMR, as well as 31P-NMR, fast-atom-bombardment mass spectrometry (FAB MS) and FAB MS/MS in positive and negative modes. The isolated core heptasaccharide (OS) was composed of 2 units D-glucose, 3 units LD-heptose and 1 unit each of DD-heptose and 3-deoxy-D-manno-octulosonic acid. Methylation analysis indicated that OS was highly branched with terminal location of the two glucoses and the DD-heptose unit, which was partially (to about 40%) phosphorylated at C7. These combined studies allowed us to formulate the structure of the inner core region as shown in Scheme 1. The substitution of the 7-position of the terminally linked DD-heptose unit by phosphate could be recognized by MS characterization of permethylated DD-heptose-7-phosphate (alditol acetate) and the extent of the substitution by the ratio of the two well separated 1H signals of DD-heptose in 500-MHz 1H-NMR. Negative FAB MS of OS also indicated the presence of smaller amounts of two hexasaccharides, differing from OS in lacking either one terminal unit of D-glucose or of the terminal DD-heptose, and additionally of a pentasaccharide lacking two heptosyl units, namely the terminal DD-heptose and and the subterminal LD-heptose. The presence of the smaller oligosaccharides in the OS fraction was also recognized by the methylation analysis.
In order to gain insight into the structure of human parathyroid hormone (hPTH), four fragments [hPTH(1-34), hPTH(18-48), hPTH(28-48), and hPTH(53-84)], which cover all regions of the intact hormone, have been investigated by CD and NMR spectroscopy in combination with distance geometry, and restrained molecular dynamics and energy minimization calculations, under a variety of solution conditions. Significantly, all fragments showed little propensity to form stable structures in aqueous solution alone, and it was only on the addition of trifluoroethanol (TFE) that defined structural features were observed. In an extension of earlier work [Klaus et al. (1991) Biochemistry 30, 6936-6942], hPTH(1-34) in 70% trifluoroethanol (TFE) showed two helices that were longer than in 10% TFE, but essentially showed the same characteristics. Although overlap in the 1H NMR spectra prevented the determination of quantitative NOE data for residues 26-30, the combination of the alpha-proton chemical shift data and quantitative NOE data indicated the helices extend from residues 3 to 13 and 15 to 29. No evidence was found for interaction of the two helical regions. The nature and extent of this second helix in the intact hormone were better defined from the data for hPTH(18-48). Under limiting solution conditions, where the fragment assumed its maximum helical content, a well-defined helix was observed between residues 21 and 38 with a possible discontinuity between Leu-28 and Gln-29. There was little evidence of any form of secondary structure between Gly-38 and the terminus of this fragment, Ser-48. In keeping with this result, the shorter fragment, hPTH(28-48), showed little evidence of stable secondary structure on addition of TFE. From the alpha-proton chemical shifts residues 23-27 appeared to sustain helical structure more readily than the rest of molecule under all solution regimes in both hPTH(1-34) and hPTH(18-48). In contrast to the other two longer fragments hPTH(53-84) showed little propensity for helical secondary structure even at the highest TFE concentrations. However, there was evidence that the molecule did adopt a defined three-dimensional structure. Various long-range NOE's were observed in 10% TFE that allowed the calculation of an open tertiary structure consisting of an initial series of turns surrounded by a loop structure of several loose turns.(ABSTRACT TRUNCATED AT 400 WORDS)