Structures of sialylated oligosaccharides of human erythropoietin expressed in recombinant BHK-21 cells

The native structures of the Asn-linked oligosaccharides and the O-glycans at Ser126 of human erythropoietin expressed from recombinant BHK cells have been elucidated. Enzymatically released N-glycans were studied by methylation analyses, fast-atom-bombardment mass spectrometry as well as one- and two-dimensional 1H-NMR spectrometry at 600 MHz. Many (82.7%) were found to be tetraantennary N-acetyllactosamine-type (22.8% with one, 3.6% with two and 0.4% with three N-acetyllactosamine repeats) being tetrasialylated (41%), trisialylated (29.6%) and disialylated (12.2%). A few (9.7%; 4.1% 2,4-branched, 5.6%, 2,6-branched) of the chains were triantennary (5.4% trisialyl, 4.3% disialyl) and 4.6% were of the disialyl diantennary type. Almost all of the innermost GlcNAc residues were alpha 1-6 fucosylated and NeuAc was exclusively alpha 2-3 linked to Gal beta 1-4GlcNAc-R; 60% of the protein was found to be O-glycosylated at Ser126; structures were monosialylated (70%) or disialylated (30%) forms of the Gal beta 1-3GalNAc core type. Glycosylation patterns at individual Asn-Xaa-Thr/Ser sites were determined by analytical high-pH anion-exchange chromatography with pulsed amperometric detection. Only tetraantennary chains with 0-3 N-acetyllactosamine repeats were detected at Asn38 and Asn83, while almost all of the di- and triantennary oligosaccharides were attached to Asn24. Batch analysis of different preparations of recombinant erythropoietin revealed the high reproducibility of the production procedure. Structures containing terminal GalNAc-GlcNAc were detected in small amounts in a few batches.     

A comparative evaluation of two sensitive serum neutralization tests for bovine herpesvirus-1 antibodies

The need for frequent injections and monitoring, the possibility of multiple gestations, and the higher cost compared to clomiphene citrate, prevents many clinicians from using human menopausal gonadotrophin (HMG) for ovulation induction. A sequential medication regimen, in which HMG is taken after clomiphene, overcomes these problems. We retrospectively compared per cycle fecundity and birth rates in 119 cycles of clomiphene-HMG, 524 cycles of clomiphene alone, 57 cycles of HMG alone, and 79 cycles of concurrent HMG and clomiphene in patients receiving intra-uterine insemination (IUI), who were free of endometriosis or tubal disease. Per cycle fecundity for clomiphene-HMG was 22% [95% confidence interval (CI) 12-34%], double that of clomiphene alone (11%) (95% CI 8-14%) (P < 0.01), and equal to HMG alone (18%) (95% CI 7-29%) or HMG and clomiphene together (19%) (95% CI 10-28%). The multiple birth rate for clomiphene-HMG (7/21) equalled that for HMG alone (3/12) and HMG and clomiphene together (3/8). The average number of ampoules of HMG required [follicle stimulating hormone (FSH) 75 mIU, luteinizing hormone (LH) 75 mIU] was decreased by 65% from 24.5 +/- 1.0 for HMG or HMG and clomiphene together to 8.6 +/- 0.3 for clomiphene-HMG (P < 0.001). Per cycle fecundity was identical when one, two or three ampoules of HMG per day were administered after clomiphene. We conclude that ovulation induction with sequential clomiphene-HMG results in fecundity double that of clomiphene alone and equal to HMG alone or concurrent with clomiphene, thereby reducing the requirement for HMG.     

Border crossing for hospital care and its implications for the use of statewide data

A major concern of researchers using state data sets for population-based analyses and market share studies in the health care sector is the potential bias caused by 'border crossing'--patients receiving care out of state. By using the Health Care Financing Administration (HCFA) discharge abstract files for 1987 and 1988, we found that 'border crossing' is not a serious problem for the two large states we examined. Only 4.4% of New York patients and 2.15% of California patients received care out of state. At the county and zip code level, 'border crossing' is more frequent but tends to be concentrated in areas adjacent to other states. Even excluding all zips with more than 10% of patients crossing the 'border' results in a small loss of patients (2.2% for New York and 1.0% for California).     

Annealing behavior of Ar ion irradiated Bi2Sr2Can-1CunO4+2n superlattice and intergrowth films

Artificially layered Bi₂Sr₂Ca_n–1Cu_nO_4+n films were synthesized by sequential sputter deposition of BiO, SrCu_0.5O_1.5 and CaCuO₂ layers. Annealing behavior of these films which were irradiated by Ar ions was studied. Defect assisted improvement of their crystalline perfection is expected which might results in the improvement of the superconducting properties of these films. An artificial film, such as an intergrowth of 2223 and 2234 phases, and superstructure films of (2245)₁(2201)₁ and (2234)₁(2212)₁, were irradiated by Ar ions (150 keV, 2–10×10¹² ions/cm₂) and annealed at 730 °C. An improvement of superconducting transition temperatures were observed.     

Prediction of fatigue screw loosening in anterior spinal fixation using dual energy x-ray absorptiometry

STUDY DESIGN: A biomechanical study was performed to investigate a relation between the bone mineral density of the vertebral body and the number of loading cycles to induce fatigue loosening of an anterior vertebral screw. OBJECTIVES: The objective of this study was to investigate the potential usefulness of dual energy x-ray absorptiometry of measuring bone mineral density of the vertebral body in predicting the fatigue loosening of th anterior vertebral screw. SUMMARY OF BACKGROUND DATA: Loosening of the vertebral body screw is a well know failure in spinal instrumentation, and more commonly observed than pullout failure. The relation between bone mineral density and pullout strength of the screw has been investigated previously, but no studies are available on the fatigue loosening in anterior spinal fixation. METHODS: Bone mineral density was measured using dual energy x-ray absorptiometry and the screw loosening was produce by a cyclic loading in the cephalad-caudal direction. Screw loosening was defined as 1 mm displacement of the screw relative to bone, and the number of loading cycles to induce the screw loosening was obtained and statistically correlated with bone mineral density. RESULTS: There was a positive correlation between the number of loading cycles to induce screw loosening and bone mineral density (R = 0.8, P < 0.01). The average number of loading cycles to induce screw loosening was significantly less for specimens with bone mineral density < 0.45 g/cm2 compared to those with bone mineral density > or = g/cm2. CONCLUSIONS: These findings suggest that bone mineral density may be a good predictor of anterior vertebral screw loosening. Bone mineral density < 0.45 g/cm2 may be critical value of loosening of the anterior vertebral body screw. However, further biomechanical and clinical studies are required before using threshold value clinically.     

Development of the Base Station Controller and Manager in the CDMA Mobile System

The base station (BS) in the CDMA Mobile System (CMS) connects calls through the radio interface and is designed to provide mobile subscribers with high quality service in spite of mobile subscribers’ motions. The BS consists of multiple base station transceiver subsystems (BTSs), a base station controller (BSC) and a base station manager (BSM). This paper is concerned with the BSC and the BSM. The BSC is located between the BTSs and the mobile switching center (MSC) connected with the public network, and is responsible for controlling mobile calls from and to mobile subscribers via the BTSs. The BSM provides operator-interfaces per the BS and takes responsibility of operation and maintenance (OAM) of the BS. Design of the BSC is based on two module types: functional module and unit module. The functional module is used to support new services easily and the unit module to increase the system capacity economically. Both modular types are easily achieved by inserting the corresponding modules to the system. Particularly, in order to efficiently support the soft handover which is one of CDMA superior advantages, the BSC adopts a large high-speed packet switch connecting up to 512 BTSs, and thus mobile subscribers can be provided with soft handover in high probability. The BSM is based on a commercial workstation to support OAM functions efficiently and guarantee high reliability of the functions. The BSM uses graphical user interface (GUI) for efficient OAM functions of the BS.     

Precision and accuracy of total body bone mass and body composition measurements in the rat using x-ray-based dual photon absorptiometry

Lean body mass (LBM), total body bone mineral mass (BMC), total body bone areal density (BMD), and body fat mass (FM) were measured in rats by dual photon absorptiometry (DXA), using two different instruments. The coefficients of variation for repeated measurements of LBM and FM were about 0.4 and 2.5%, respectively, over an animal body weight range of 150 to 600 g. For BMC and BMD, the coefficients of variation were less than 2%. The correlation coefficients for LBM, FM, BMC, and BMD measured on the two densitometers were all greater than 0.94. The slope of the regression line relating LBM measured by DXA and LBM measured by carcass analysis was 0.999, and the correlation coefficient was 0.99. For FM the slope was 1.05, and the correlation coefficient was 0.98. BMC measures by DXA were falsely low in small animals. For larger animals, the correlation between BMC and ash weight was 0.93, but the slope of the regression line was 0.78. DXA measures of LBM and FM were accurate and reproducible for rats weighing between 150 and 600 g. There was a size-dependent error in BMC, which will be significant in longitudinal measurements of bone mass.     

Substrate transport and cocaine binding of human dopamine transporter is reduced by substitution of carboxyl tail with that of bovine dopamine transporter

A chimeric dopamine transporter (DAT) cDNA encoding mutant human DAT (hDAT) protein in which the intracellular carboxyl-terminal tail is replaced by that of the bovine dopamine transporter (bDAT) was constructed. The chimeric hDAT cDNA was expressed in COS-7 cells, and [3H]dopamine and [3H]MPP+ uptake and [3H]CFT binding capacities were assessed. Substrate transport and ligand binding of bDAT were reduced by 32-43% as a result of substitution of the carboxyl tail in hDAT, suggesting that the functional characteristics of bDAT arise from differences in the carboxyl tail between human and bovine DAT. Thus, it appears that the sequences encoded within the carboxyl terminal of DAT would be one of the important determinants for its functions.     

Regulation of tyrosine hydroxylase and aromatic L-amino acid decarboxylase by dopaminergic drugs

We provide evidence that dopamine receptors differentially modulate tyrosine hydroxylase and aromatic L-amino acid decarboxylase in the mouse striatum. The dopamine D1 receptor family (D1-like) antagonist, R(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1 H-3-benazepine (SCH 23390), elevated aromatic L-amino acid decarboxylase activity and protein content in striatum, as well as the mRNA for the enzyme in midbrain. The dopamine D1-like receptor agonist, (+/-)-1-phenyl-2,3,4,5-tetrahydro-(1 H)-3-benzazepine-7,8-diol (SKF 38393), had no effect on aromatic L-amino acid decarboxylase. The dopamine D1-like drugs had no effect on tyrosine hydroxylase. In contrast, the dopamine D2 receptor family (D2-like) antagonists haloperidol and spiperone elevated both tyrosine hydroxylase and aromatic L-amino acid decarboxylase activities. The increase in aromatic L-amino acid decarboxylase activity was accompanied by elevated enzyme protein content but not mRNA. The dopamine D2-like receptor agonists, bromocriptine, quinpirole and (+/-)-7-hydroxydipropylaminotetralin (7-OH-DPAT), all decreased striatal tyrosine hydroxylase. Under the conditions used, bromocriptine and 7-OH-DPAT, but not quinpirole, decreased aromatic L-amino acid decarboxylase activity of striatum. Both the dopamine D1- and D2-like receptor antagonists enhanced the turnover of striatal dopamine to differing degrees, as judged by the ratio of acid metabolites of dopamine to dopamine. Taken together our results indicate that aromatic L-amino acid decarboxylase can be modulated independently of tyrosine hydroxylase.