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101.
JE Manning DN Batson TW Gansman CA Murphy SG Perretta EA Norfleet 《Canadian Metallurgical Quarterly》1997,4(9):883-890
With the aim of developing foetal gene therapy for cystic fibrosis, we have investigated the possibility of gene targeting to the mouse foetus with two different viral vector systems and at different times of gestation. We report here that recombinant retrovirus producing cells administered into the intra-amniotic cavity of mid- to late-gestation mouse MF1 foetuses survive in the amniotic fluid and are able to engraft to a certain extent in foetal tissues. By production of infectious virus they mediate transduction and beta-galactosidase transgene expression in neighbouring foetal tissues 24 to 72 h following injection. Retrovirus producer cells could, therefore, become a means to overcome the limitations of low retroviral titre, for in vivo foetal gene transfer. To investigate the developmental stage at which transduction of the airways and enteral systems can be obtained we also administered a highly infective first generation adenoviral vector (AdRSV beta gal) into the amniotic cavity of foetal mice between 13 to 16 days post coitus, beta-galactosidase activity was detected between 24 to 120 h after injection. The highest levels of transgene expression were generally observed between 48 to 72 h following injection of the adenoviral vector. We demonstrate that infection of the pulmonary airways is dependent on the developmental stage of the foetus and can be achieved on the 15th day of gestation. 相似文献
102.
Immunization by peptides based on the repeat sequences of Plasmodium falciparum or P. vivax antigen(s) have shown inconsistent results during clinical trials in humans. This could be attributed to the lack of T-cell help or antigenic polymorphism. Thus, attention has been focused towards the more conserved non-repeat regions. The present study was undertaken to map the antigenic determinant in the vicinity of region II (outside the repeat) of CS protein of P. vivax. The immunogenicity of the peptide was studied alone and after linking with polytuftsin (PT), using alum and Freund's adjuvant, in inbred strains of mice with different genetic backgrounds. The humoral response and antigen induced T-cell proliferation assays clearly demonstrated the immunomodulatory activity of PT. Comparable results were observed with antigen(s) administered either in alum or Freund's adjuvant. The induction of IgG2a and IgG2b antibody isotypes by both, peptide as well as the conjugate, may indicate that the T-helper response involved is of Th1 type. Further the immunofluorescence studies have shown that antibodies recognized the air dried sporozoites of P. cynomolgi. The results thus show that the above sequence has overlapping B and T-cell determinants and that alum can be substituted for Freund's adjuvant in generating an effective immune response. 相似文献
103.
DN Angelov M Walther M Streppel O Guntinas-Lichius AM van Dam E Stennert WF Neiss 《Canadian Metallurgical Quarterly》1998,54(6):820-827
An adeno-associated virus (AAV) vector, expressing genes for human tyrosine hydroxylase (TH) and aromatic amino acid decarboxylase (AADC), demonstrated significantly increased production of dopamine in 293 (human embryonic kidney) cells. This bicistronic vector was used to transduce striatal cells of six asymptomatic but dopamine-depleted monkeys which had been treated with the neurotoxin MPTP. Striatal cells were immunoreactive for the vector-encoded TH after stereotactic injection for periods up to 134 days, with biochemical effects consistent with dopamine biosynthetic enzyme expression. A subsequent experiment was carried out in six more severely depleted and parkinsonian monkeys. Several TH/aadc-treated monkeys showed elevated levels of dopamine near injection tracts after 2.5 months. Two monkeys that received a beta-galactosidase expressing vector showed no change in striatal dopamine. Behavioral changes could not be statistically related to the vector treatment groups. Toxicity was limited to transient fever in several animals and severe hyperactivity in one animal in the first days after injection with no associated histological evidence of inflammation. This study shows the successful transfection of primate neurons over a period up to 2.5 months with suggestive evidence of biochemical phenotypic effects and without significant toxicity. While supporting the idea of an in vivo gene therapy for Parkinson's disease, more consistent and longer lasting biochemical and behavioral effects will be necessary to establish the feasibility of this appraoch in a primate model of parkinsonism. 相似文献
104.
E Roine DM Raineri M Romantschuk M Wilson DN Nunn 《Canadian Metallurgical Quarterly》1998,11(11):1048-1056
Many strains of Pseudomonas syringae produce retractile pili that act as receptors for lytic bacteriophage phi 6. As these are also characteristics of type IV pili, it was postulated that P. syringae may possess genes for type IV pilus biogenesis. A cosmid clone bank of P. syringae pv. tomato DC3000 genomic DNA was used to complement a mutant of Pseudomonas aeruginosa defective in the PilD (XcpA) prepilin peptidase gene by selection for restoration of extracellular protein secretion, a function also known to require PilD. A cosmid able to complement this mutant was also able to complement mutations in the pilB and pilC genes, suggesting that, if the organization of these genes is similar to that of P. aeruginosa, the cosmid may contain the P. syringae pilA. This was confirmed by sequencing a region from this plasmid that was shown to hybridize at low stringency to the P. aeruginosa pilA gene. The deduced P. syringae PilA polypeptide possesses the characteristic properties of the type IV pilins. Heterologous expression of the P. syringae pilA in P. aeruginosa was also shown, conferring not only phi 6 phage sensitivity to P. aeruginosa pilA mutants but also sensitivity to PO4, a lytic bacteriophage specific for the pilus of P. aeruginosa. This suggests that additional components might be present in the mature pilus of P. aeruginosa that are the true receptors for this phage. Chromosomal mutations in P. syringae pv. tomato DC3000 pilA and pilD genes were shown to abolish its sensitivity to bacteriophage phi 6. To determine the importance of P. syringae pilus in plant leaf interactions, these mutations were tested under laboratory and field conditions. Although little effect was seen on pathogenicity, culturable leaf-associated population sizes of the pilA mutant were significantly different from those of the wild-type parent. In addition, the expression of the DC3000 pilA gene appears to contribute to the UV tolerance of P. syringae and may play a role in survival on the plant leaf surface. 相似文献
105.
Choline acetyltransferase catalyzes the synthesis of acetylcholine from choline and acetylcoenzyme A (ACoA) in both nervous and non-nervous tissues. Carnitine acetyltransferase occurs in several tissues and transfers acetyl groups from ACoA to carnitine forming acetylcarnitine and exhibits weak choline acetyltransferase activity. Several haloacetylcholines and haloacetylcarnitines were synthesized to develop selective inhibitors of choline acetyltransferase and carnitine acetyltransferase. Acetylcholine is a transmitter for some presynaptic neurons and/or amacrine cells in retina. Selective inhibitors of choline acetyltransferase and carnitine acetyltransferase were used in the evaluation of choline acetyltransferase and carnitine acetyltransferase activities in the rat retina. Choline acetyltransferase and carnitine acetyltransferase activities were assayed by transferring of [14C]acetyl group from [14C]ACoA to choline or carnitine and estimating [14C]-acetylcholine or [14C]acetylcarnitine. This study gave the following results: (a) Bromoacetylcholine (BrACh) was a selective inhibitor of purified choline acetyltransferase (I50, 2.2 microM); (b) (R)-bromoacetylcarnitine [(R)-BrACa] was more potent for inhibiting purified carnitine acetyltransferase (I50, 4 microM) than purified choline acetyltransferase (I50, 46 microM); (c) Rat retinal sonicate gave choline acetyltransferase activity of 98 +/- 6 nmol of ACh formed/mg/10 min. When the carnitine acetyltransferase was completely inhibited by (R)-BrACa, the activity for choline acetyltransferase decreased to 47 +/- 1 nmol, and this decrease was possibly due to the formation of some [14C]acetylcholine by carnitine acetyltransferase. The net retinal choline acetyltransferase activity was 51 nmol acetylcholine/mg protein/10 min; (d) Rat retinal sonicate contained carnitine acetyltransferase activity of 102 +/- 7 nmol acetylcarnitine formed/mg protein/10 min. This was not altered by inhibition of choline acetyltransferase with BrACh. This means that choline acetyltransferase did not use carnitine as a substrate. Choline acetyltransferase and carnitine acetyltransferase activities did not change after dialysis of retinal sonicates at 4 degrees C for 24 hrs. These observations suggest that BrACh and (R)-BrACa are useful for assessing the correct values for choline acetyltransferase and carnitine acetyltransferase activities in retinal tissues. 相似文献
106.
W. O. Soboyejo K. T. Venkateswara Rao S. M. L. Sastry R. O. Ritchie 《Metallurgical and Materials Transactions A》1993,24(3):585-600
The results of recent studies on the fatigue and fracture behavior of extruded Ti-48A1 + 20 vol pct TiNb and hot-isostatically
pressed (“hipped”) MoSi2 + 20 vol pct Nb are presented (compositions in atomic percent unless stated otherwise). The effects of ductile phase reinforcement
of Ti-48A1 and MoSi2 on the micromechanisms of fracture under monotonie and cyclic loading are elucidated. Micromechanics models are applied to
the prediction of crack-tip shielding components, and the effects of temperature on tensile/compressive/flexure strengths
are discussed. Ductile phase toughening under monotonie loading conditions is shown to be associated with lower fatigue crack
growth resistance. The lower fatigue resistance is attributed to the absence of crack-tip shielding, higher crack opening
displacements, and the effects of inelastic strains that are developed in ductile phase-reinforced composites under cyclic
loading conditions.
S.M.L. SASTRY, formerly Program Manager and Fellow, McDonnell Douglas Research Laboratories.
This article is based on a presentation made in the symposium “Quasi-Brittle Fracture” presented during the TMS fall meeting,
Cincinnati, OH, October 21–24, 1991, under the auspices of the TMS Mechanical Metallurgy Committee and the ASM/MSD Flow and
Fracture Committee. 相似文献
107.
RP Mlcak MH Desai E Robinson RL McCauley J Richardson DN Herndon 《Canadian Metallurgical Quarterly》1995,21(5):337-339
During a transjugular intrahepatic portosystemic shunt (TIPS) procedure, a Strecker stent was accidently pushed into the superior mesenteric vein. After successful shunt placement, the stent was withdrawn into the hepatic vein. A multipurpose basket catheter was attached to the distal end of the stent and a loop snare to the proximal end. In this way it was possible to stretch the stent and retrieve it percutaneously through the jugular sheath. 相似文献
108.
S. Hariprasad D.Sc. S. M. L. Sastry Ph.D. 《JOM Journal of the Minerals, Metals and Materials Society》1995,47(10):56-59
A validated mathematical model was used to generate processing maps for producing near-net shapes of high-performance products by gas atomization and spray deposition. The processing maps are useful to optimize and control the manufacturing process on the shop floor for producing quality products. The number of trial runs and cost of production can be significantly reduced with the help of processing maps. Using a dispersion-strengthened aluminum alloy as an example, this article examines how processing maps can be used to choose a set of process variables for producing the end product of desired quality. 相似文献
109.
The macula is a constituent of the sensory retina that is necessary for sharp contrast and color vision. A significant relationship has been found between tobacco smoking and age-related macular degeneration. Opsin, rhodopsin and phospholipase A2 (PLA2) are located in excitable membranes of retina which are enriched with polyunsaturated fatty acids (PUFA). A question may arise as to whether nicotine and its major metabolite cotinine influence PLA2 so that arachidonic acid (AA) and proinflammatory prostaglandins (PG) are produced in the retina. Therefore, the effects of nicotine and cotinine on the retinal PLA2 were studied. PLA2 activity of rat retinal sonicates was assayed using 1-palmitoyl-2[1-14C]arachidonyl-Phosphatidylethanolamine (PE, 2.2 nmol) as a substrate in Tris buffer (10 mM, pH 7.4) at 37 degrees C with and without nicotine or cotinine in the assay medium. These studies gave the following results: (1) Rat retinal PLA2 activity was 4.2+/-0.8 pmol PE hydrolyzed/100 ng protein/hr. (2) Nicotine in low concentrations (1-150 nM) activated PLA2 (EC50 5 nM). (3) Cotinine also activated PLA2 (EC50 300 nM). (4) Only high concentrations of nicotine (> 1.0 microM) and cotinine (> 25 microM) exhibit inhibition of PLA2. (5) All three known PLA2 inhibitors, mepacrine, 4-bromophenacyl bromide and bromoacetylcholine bromide (IC50: 0.5mM, 88 microM, 30 mM, respectively) inhibited retinal PLA2 activity. These observations suggest that polyunsaturated fatty acids are cleaved, and arachidonic acid, the precursor for prostaglandins and related pro-inflammatory mediators, is liberated by nicotine and cotinine. Oxidative stress (reduced levels of antioxidants), vascular insufficiency, as well as activation of PLA2 by nicotine and cotinine may contribute for retinal degeneration in smokers during aging. 相似文献
110.
SM Palmer NG Henshaw DN Howell SE Miller RD Davis VF Tapson 《Canadian Metallurgical Quarterly》1998,113(4):944-950
STUDY OBJECTIVE: To define the epidemiology, clinical manifestations, and long-term complications of respiratory viral infections in adult lung transplant recipients. DESIGN: Retrospective review of the records of 122 adult lung transplant recipients over a 5-year period at one institution. RESULTS: Ten episodes of infection with respiratory syncytial virus, parainfluenza, influenza, or adenovirus were identified. All patients presented with symptoms of respiratory tract infection. Two patients died acutely and four patients subsequently had development of obliterative bronchiolitis (OB). CONCLUSIONS: These data suggest community respiratory viral infections cause significant morbidity and mortality in lung transplant recipients. Further prospective studies are warranted to clarify the relationship between respiratory viral infection and OB and to define the optimal therapy for these viral infections. 相似文献