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141.
Analysis of lipidation of recombinant Lyme vaccine protein (rOspA) by electrospray mass spectroscopy
XD Wu KL Marakov PA Martinez PH McVerry DA Malinzak 《Canadian Metallurgical Quarterly》1997,16(4):613-617
Lipidation at the N-terminal of recombinant outer surface protein A (rOspA) is an important determinant of its immunogenicity. Lipidation patterns of rOspA can be sensitive to processing environments and storage conditions. In order to assure product consistency and stability. it is essential to characterize and monitor lipidation patterns of rOspA through its life-cycle. Electrospray mass spectroscopy combined with maximum entropy calculation was employed to analyze the lipidation of rOspA. The results revealed that more than 90% of protein is a tri-lipidated rOspA and the remainder is di-lipidated. It was demonstrated that the method is both sensitive and quantitative and has the potential to be used for routine quality control and stability testing. 相似文献
142.
Vitamin K-dependent carboxylase catalyzes the post-translational gamma-carboxylation of 9-12 glutamyl residues of several blood coagulation proteins. Carboxylase purified from Chinese hamster ovary (CHO) cells as a recombinant FLAG-carboxylase fusion protein [Sugiura, I., et al. (1996) J. Biol. Chem. 271, 17837-17844] was utilized with pentapeptide substrate FL[3H-R,S]EAL with high specific radioactivity to probe the timing of glutamyl Cgamma-3H cleavage relative to Cgamma-COO- bond formation by 14CO2 incorporation rates. Studies were conducted over a range of NaH14CO3 concentrations to assess uncoupling of gamma-glutamyl carbanion formation and over a range of concentrations of ProPT18, the 18-residue peptide corresponding to the -18 to -1 propeptide region of prothrombin known to affect the catalytic efficiency of carboxylase. At saturation, ProPT18 accelerates Cgamma-3H cleavage 11-13-fold and Cgamma-14CO2- formation 6-7-fold, converting a Cgamma-3H cleavage/Cgamma-14CO2- formation ratio of 1.2-1.4 in the absence of ProPT18 to 2.3-2.8 in its presence, a relative increase in and uncoupling of Cgamma-3H cleavage from C-C bond formation. When the HCO3- concentration was varied, the V/K3H+/V/K14CO2 ratios rose as HCO3- fractional saturation dropped to a ratio of 9.3-10.8/l at low bicarbonate, indicating an uncoupling of nine out of ten gamma-glutamyl carbanion formations from carboxylative capture, consistent with prior reports on microsomal enzyme [Larson, A. E., et al. (1981) J. Biol. Chem. 256, 11032-11035]. These results with pentapeptide substrate FLEAL validate reversible gamma-glutamyl carbanion formation by pure carboxylase and indicate the ProPT18 increase in catalytic efficiency is in selective lowering of an energy barrier preceding the gamma-glutamyl carbanion intermediate. 相似文献
143.
KL Manchester 《Canadian Metallurgical Quarterly》1997,230(1):27-29
Acute idiopathic thrombocytopaenic purpura is the most common of the thrombocytopaenias of childhood. Clinically it is associated with petechiae, mucocutaneous bleeding and occasionally haemorrhage into tissues. The oral mucosa is frequently involved. This paper describes a case presenting in general dental practice. 相似文献
144.
P Devita T Hortobagyi J Barrier M Torry KL Glover DL Speroni J Money MT Mahar 《Canadian Metallurgical Quarterly》1997,29(7):853-859
Gait analyses of rehabilitated individuals with anterior cruciate ligament (ACL) deficiency and reconstruction have identified the final adaptations of increased hip extensor torque and hamstring electromyography (EMG) and decreased knee extensor torque and quadriceps EMG during stance. The initial adaptations to injury and surgery are, however, unknown as are the factors that influence the development of the adaptations. Identification of the initial response to injury would provide a basis for determining whether the final adaptations are learned automatically or if they are the result of a lengthy training period in which various factors may affect their development. The purpose of the study was to evaluate the initial effects of ACL injury and reconstruction surgery on joint kinematics, kinetics, and energetics, during walking. Injured limbs from nine subjects with ACL injury were tested 2 wk after injury, and 3 and 5 wk after surgery. Ten healthy subjects were tested. Kinematic and ground reaction data were collected and combined with inverse dynamics to calculate the joint torques and powers. A knee extensor torque throughout most of stance was observed in the injured limbs at all test sessions. This result was in conflict with previous observations of reduced extensor torque or a flexor torque in rehabilitated patients with ACL reconstruction and patients with ACL deficiency. This result also differed from the typical midstance extensor then flexor torque in healthy control subjects. Trend analysis showed a significant (P < 0.001) change in average position at the hip and knee, extensor angular impulse at the hip, and positive work done at the hip 3 wk after surgery followed by a partial rehabilitation at 5 wk after surgery. Power and work produced at the knee were reduced fivefold (P < 0.001) after 5 wk of rehabilitation and did not recover to pre-surgical levels. The existence of a long-lasting knee extensor torque 2 wk after injury indicated that the adaptation process to ACL deficiency is lengthy, requiring many gait cycles, and that numerous factors could be involved in learning the adaptations. 相似文献
145.
The data collected by a postal questionnaire sent to 3772 randomly selected dairy farmers in England and the border regions in Wales were used to estimate the relationships between the presence of clinical Johne's disease and farm and management factors associated with that disease. Two binary outcomes (case reported in 1993, case reported in 1994) and 27 predictor variables were considered. Only two variables were consistently and significantly associated with clinical disease in multivariable analysis. Farms on which Channel Island breeds were predominant were associated with an increased risk of reporting disease (odds ratios (ORs) ranged from 10.9 to 12.9). The presence of farmed deer on the farm also increased the risk of reporting disease (ORs ranged from 15.2 to 209.3). There were other significant but inconsistent associations involving the source of replacements, age of first-offering hay, type of concentrate feed to calves, and calving in individual pens when the cows were at grass. Since Johne's disease is predominantly subclinical, these contributing factors may play important roles in switching subclinical infection to overt disease. 相似文献
146.
Fluconazole is a triazole antifungal agent which is now an established part of therapy in patients with immune deficiencies. It is effective against oropharyngeal/oesophageal candidiasis (candidosis) when used orally once daily either as treatment or secondary prophylaxis in patients with AIDS or as treatment or primary prophylaxis in neutropenia associated with cancer therapy. Fluconazole also resolves symptoms in up to 60% of patients with cryptococcal meningitis and AIDS. However, in this infection its efficacy as treatment relative to that of amphotericin B is equivocal, and its major role is as the drug of choice for maintenance therapy following amphotericin B induction. In this regard, fluconazole has been proven superior to amphotericin B and to itraconazole 200 mg/day. Comparisons with other drugs used for the treatment of mucosal candidiasis in patients with AIDS show fluconazole to be superior to nystatin, similar to itraconazole and at least as effective as clotrimazole and ketoconazole; it was more so than the latter azole in 1 study. In patients undergoing chemotherapy or bone marrow transplantation, fluconazole as primary prophylaxis has produced greater clinical benefit than a clotrimazole regimen. The incidence of adverse events appears to be somewhat higher in patients with AIDS compared with HIV-negative cohorts, but the qualitative pattern of events is similar. The most frequent events are gastrointestinal complaints, headache and skin rash: rare exfoliative skin reactions and isolated instances of clinically overt hepatic dysfunction have occurred in patients with AIDS. Issues yet to be clarified include: the use of fluconazole in children with AIDS, in whom results have been promising; its efficacy against other fungal infections encountered in immunocompromised patients; whether the drug influences mortality, as has been suggested by one placebo-controlled trial in patients undergoing bone marrow transplant; and the appropriateness of its potential for use as primary prophylaxis against cryptococcal meningitis in patients with AIDS, where it shows efficacy but there is concern over increasing risk of development of secondary resistance. Notwithstanding these undefined aspects of its clinical profile, fluconazole is now confirmed as an important antifungal drug in the management of fungal infections in patients with immune deficiencies. In patients with AIDS it is the present drug of choice as maintenance therapy against cryptococcal meningitis and is a preferred agent for secondary prophylaxis against candidal infections; it is also a favoured agent for primary prophylaxis in patients at risk because of neutropenia associated with chemotherapy or bone marrow transplantation . 相似文献
147.
148.
Laser Doppler flowmetry was used intraoperatively to monitor femoral head perfusion during open reduction of 13 congenital hip dislocations in 11 patients. Laser Doppler determinations ranged from 12 to 400 mV before reduction and 30 to 300 mV after reduction. Three patients had magnitude changes in excess of 50%. One had increased perfusion, and two had decreased blood flow. Avascular necrosis of the hip occurred in one patient that was not predicted by laser Doppler flowmetry. Femoral head perfusion measured 175 mV for the dislocated hip and 180 mV after reduction of the femoral head and completion of the pelvic osteotomy. The authors conclude that laser Doppler flowmetry is not a reliable method for monitoring femoral head perfusion during open reduction of the hip for developmental hip dysplasia. 相似文献
149.
BA Bart LK Shaw CB McCants DF Fortin KL Lee RM Califf CM O'Connor 《Canadian Metallurgical Quarterly》1997,30(4):1002-1008
OBJECTIVES: We sought to characterize the clinical determinants of mortality in patients with angiographically diagnosed ischemic or nonischemic cardiomyopathy. BACKGROUND: Patients with ischemic cardiomyopathy may have a worse prognosis than patients with nonischemic cardiomyopathy. Few studies have assessed the effect of ischemic versus nonischemic etiology on outcomes. METHODS: We analyzed prospectively collected data on 3,787 patients with a left ventricular ejection fraction < or = 40% who underwent coronary angiography. Patients were considered to have ischemic cardiomyopathy (n = 3,112) if they had a history of myocardial infarction, percutaneous transluminal coronary angioplasty, coronary artery bypass graft surgery or at least one major epicardial coronary artery with > or = 75% stenosis; all others were considered to have nonischemic cardiomyopathy (n = 675). RESULTS: The median age, ejection fraction and proportion of patients with New York Heart Association functional class III or IV symptoms for the nonischemic and ischemic groups were 55 years versus 63 years, 27% versus 32% and 57% versus 25%, respectively. After adjustment for baseline clinical risk factors and presenting characteristics, ischemic etiology remained an important independent predictor of 5-year mortality (p < 0.0001). The extent of coronary artery disease was a better predictor of survival than ischemic or nonischemic etiology (log likelihood chi-square 700 vs. 675, respectively). CONCLUSIONS: Ischemic etiology is a significant independent predictor of mortality in patients with cardiomyopathy. However, the extent of coronary artery disease contributes more prognostic information than the clinical diagnosis of ischemic or nonischemic cardiomyopathy. Further research is needed to refine the clinical definition of ischemic cardiomyopathy so that physicians can appropriately prescribe treatment and accurately predict outcome. 相似文献
150.
Leptin is present in human milk and is related to maternal plasma leptin concentration and adiposity
KL Houseknecht MK McGuire CP Portocarrero MA McGuire K Beerman 《Canadian Metallurgical Quarterly》1997,240(3):742-747
Leptin is elevated during pregnancy and may be involved in the regulation of milk production in women. Immunoreactive leptin was quantified in human milk by modified radioimmunoassay. Leptin concentration was higher in whole vs. skim milk fractions; however, leptin concentration was not correlated with percentage milk fat. Leptin concentrations in whole and skim milk were correlated with maternal plasma leptin concentrations, maternal body weight, body mass index, and tricep skinfold thickness, but not with plasma insulin concentration. These data provide the first evidence for the presence of leptin in human milk in the range of concentrations found in human plasma and indicate that the concentration of leptin in milk reflects maternal adiposity. Determining the biological role(s) of milk-borne leptin could add to our understanding of neonatal metabolism and the mechanisms underlying the development of body fat and obesity in humans. 相似文献