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At least 541 records · Page 30Linked to original sources
PubMed1999

A solid-phase immobilized epitope immunoassay (SPIE-IA) permitting very sensitive and specific measurement of angiotensin II in plasma.

We have developed a new enzyme immunometric assay for angiotensin II (AII) based on SPIE-IA technology (solid-phase immobilized epitope-immunoassay). A monoclonal antibody with optimal properties (mAb3 131) was selected from a series of 19 anti-AII mAbs. The mAb had to be purified from ascitic fluid in a specific manner in order to remove endogenous AII from the antibody-binding sites. We established a sensitive (minimum detectable concentration 0.5 pg/ml) and precise (CV below 15% in the 2-100 pg/ml range) SPIE-IA. Using different AII-related peptides, we observed that this new assay has a specificity profile that compares favourably with the corresponding competitive immunoassay. We have used the assay to measure AII in 42 plasma samples, and demonstrated a good correlation with values obtained using a commercial radioimmunoassay. Assay specificity was supported by HPLC fractionation experiments, confirming the absence of interference induced by endogenous AII-related products.

Acetylcholinesterase↗
PubMed2003

A single administration of adenoviral-mediated HGF cDNA permits survival of mice from acute hepatic failure.

Heptatocyte growth factor (HGF) having a variety of biological activity was suggested as a protective agent against acute toxic hepatic injury or a potentially therapeutic agent. For the efficient in vivo application of this factor, we employed adenoviral-mediated HGF gene delivery system. In this study, we constructed E1-deleted recombinant adenovirus carrying cDNA of human HGF (Ad.hHGF) and elucidated that HGF was efficiently expressed in the liver of C57/BL mice. A mouse model of acute hepatic failure was induced by high dose (1000mg/kg) of thioacetamide (TA) administration. Mice infected with Ad.hHGF showed a dramatic resistance to TA-induced acute hepatic injury. Serum ALT was increased transiently and then the level was normalized in Ad.hHGF-infected mice with TA administration. Furthermore, the survival rate was remarkably enhanced in the mice infected with Ad.hHGF. In the histological examination, massive hepatic necrosis induced by TA was almost completely protected by HGF produced by Ad.hHGF. Our results indicate that a single dose of HGF-encoding adenoviral vector maintained liver function and prevented the progression of liver necrosis in a mouse model of acute hepatic failure.

Adenoviridae↗
PubMed1995

Correct preoperative localization: does it permit a change in operative strategy for primary hyperparathyroidism?

BACKGROUND: A meticulous bilateral neck exploration by an experienced endocrine surgeon offers a high cure rate with low morbidity for patients with primary hyperparathyroidism. The advent of localizing studies raises the possibility of unilateral neck exploration. The cost-effectiveness of preoperative localizing studies and unilateral neck exploration in primary hyperparathyroidism are controversial issues. This study was designed to determine the risks of missing a contralateral pathologic parathyroid gland in patients with preoperative localization that was confirmed at neck exploration. METHODS: Preoperative studies (ultrasonography, nuclear radioactive imaging scan, or both) were performed in 79 patients with primary hyperparathyroidism. In 58 patients a definite localization of an enlarged parathyroid gland was confirmed at operation. All patients underwent a meticulous bilateral neck exploration. RESULTS: Unilateral neck exploration was feasible only in 73.4% of the patients, according to our localizing modalities, and an additional enlarged parathyroid gland on the contralateral side, not detected before operation, was revealed in five patients (8.6%). False-positive rates were 1.7% for ultrasonography and 13% for scan. CONCLUSIONS: These results indicate an unacceptably high surgical failure rate for unilateral neck exploration guided by preoperative localizing studies compared with a bilateral neck exploration by an experienced endocrine surgeon, questioning the cost-effectiveness of preoperative localizing studies.

Adenoma↗
PubMed1997

In vivo depletion of host CD4+ and CD8+ cells permits engraftment of bone marrow stem cells and tolerance induction with minimal conditioning.

BACKGROUND: Solid organ transplantation has become the preferred approach for the treatment of end-stage organ failure. However, the toxicity associated with the nonspecific immunosuppression essential to graft survival is substantial. Bone marrow transplantation (BMT) can overcome these limitations by the induction of donor-specific tolerance. The morbidity and mortality associated with fully ablative conditioning used to achieve engraftment has prevented the clinical application of BMT for induction of tolerance for solid organ transplantation. Although it was previously believed that fully ablative conditioning was essential to achieve engraftment, it has recently become apparent that partial conditioning may be sufficient to achieve chimerism and tolerance. The focus of this study was to characterize which cells in the host microenvironment must be eliminated for engraftment of MHC-disparate bone marrow to be achieved. METHODS: C57BL/10SnJ mice were depleted of CD4+, CD8+, or both cell types with monoclonal antibodies before irradiation with 300 centigray (cGy) and transplantation of 15 x 10(6) allogeneic (B10.BR) bone marrow cells. Two days after transplantation the animals were treated with 200 mg/kg cyclophosphamide. Animals were typed for chimerism at 28 days and monthly thereafter. RESULTS: The combination of CD4+ and CD8+ depletion resulted in multilineage engraftment in 76.5% of the animals at a level of 57.1 +/- 17.7%. The depletion of CD4+ cells alone was not sufficient to allow engraftment, whereas depletion of CD8+ cells alone was. CONCLUSIONS: T cells in the recipient's marrow space play an important role in hindering allogeneic engraftment in the mouse. The T-cell subset responsible appears to be CD8+ cells rather than CD4+ cells.

Animals↗
PubMed2003

Global eddy permitting simulations of surface ocean nitrogen, iron, sulfur cycling.

The geocycles of N, Fe and S in the ocean are tightly coupled and together exert strong influence on biogeochemistry of the earth system. We investigate this interaction by inserting macro-micronutrient cycling into a high resolution ocean model that realistically represents the general circulation. Simulated chlorophyll distributions reproduce many features of satellite-based measurements of ocean color. Meridional sections through model results and seasonally averaged SeaWiFS data agree reasonably well, both in the oligotrophic gyres and along fronts. Discrepancies are associated in many cases with shelf, ridge or island effects. Dimethyl sulfide peaks and their chlorophyll correlations are similar to those obtained on major oceanographic expeditions. Lack of strong regional relationships between the sulfide and phytoplankton may be partly explained by correspondence between time constants for production and for the traversal of mesoscale transport features. In general the eddies and filaments of tropical instability waves are well represented, including the onset of the 1997-1998 El Niño. North-south shifts of the transition zone chlorophyll front appear prominently in the results.

Animals↗
PubMed1989

Cultured composite skin grafts: biological skin equivalents permitting massive expansion.

After assessment in mice, a biological skin equivalent consisting of cultured fibroblasts in a collagen gel overlain with cultured keratinocytes was applied to three patients who had had tattoos excised. In all patients the grafts took well with good cosmetic results and little or no contracture. A biopsy at 4 weeks showed that the central graft area had a fully differentiated epidermis and a mature dermis without adnexae.

Adult↗
PubMedDate not supplied

Climbing behavior permits in vivo assessment of pre- and postsynaptic extrapyramidal dopaminergic function in mice.

The natural tendency of mice to climb has been investigated in this study as an index of extrapyramidal dopaminergic function. Depending on dose, apomorphine reduced (low dose range; presynaptic dopamine receptor agonism) or increased (high dose-range; postsynaptic dopamine receptor agonism) climbing activity with respect to spontaneous basal levels of such activity in Swiss-Webster mice. We report also an increase in apomorphine-induced enhancement of vertical climbing activity in mice withdrawing from the acute effects of cesium chloride. Spontaneous climbing activity in mice could reflect dynamic extrapyramidal motor tone, upon which voluntary motor activity is superimposed and which, in humans, is adversely affected in motor disorders like parkinsonism.

Animals↗
PubMed1999

Conditions providing enhanced transfection efficiency in rat pheochromocytoma PC12 cells permit analysis of the activity of the far-upstream and proximal promoter of the brain creatine kinase gene.

While brain creatine kinase (CKB) is expressed at highest levels in the brain, where it functions in regenerating ATP, the gene elements and protein factors regulating CKB transcription in neuronal and glial cells have not been identified. To investigate the regulation of CKB in neuronal cells, we examined the expression of the promoter proximal and 5' far-upstream regions of the rat CKB gene transiently transfected into rat PC12 pheochromocytoma cells. Initially, these experiments were hampered by the extremely low transfection efficiency of PC12 cells. We increased efficiency by greater than 200-fold by employing CaPO4-precipitated DNA transfection into PC12 cells which were optimized for transient transfection by: (i) culturing cells in polylysine-coated dishes to insure attachment throughout transfection; (ii) exposing cells to transfected DNA for an optimal time and employing a glycerol shock; and, most importantly, (iii) dissociating the characteristic self-adhesive clumps of PC12 into mostly single cells. Use of the plasmid expressing green fluorescent protein allowed identification of the transfected cells that averaged 10-20% of the total. Analyses of CKB promoter-CAT gene constructs showed that in PC12 cells expression of the proximal (0.2 kb) CKB promoter was low while expression of the 1.4 kb promoter was three fold higher and the 2.9 kb promoter was ten fold higher, suggesting the presence of at least two upstream cis-acting, positive regulatory elements. In agreement, the steady-state CKB mRNA level was higher in PC12 than in other neuronal cell lines examined, possibly reflecting the effects of positive upstream factors. The results are discussed in relation to how this economical and straightforward transfection procedure may be useful in identify factors regulating the transcription of CKB and other genes expressed in neuronal cells.

Animals↗