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A. Longo M. Di Toro E. Pagani A. Carenzi 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1981,224(2):257-264
A rapid, sensitive and specific selected ion monitoring method is described for the determination of fosfomycin in plasma and urine. The extraction of the drug from serum involves deproteinization with ethanol (1 ml per 0.25 ml of serum), evaporation of an aliquot of supernatant and derivatization with a silylating mixture consisting of bistrimethylsilyl-acetamide—dichloromethane (1:1) + 5% of trimethylchlorosilane. The analysis of fosfomycin in urine requires the dilution of samples and their derivatization only. The results were compared with those obtained by analysing the same samples using a microbiological method. 相似文献
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24.
F S Jin R J Youle V G Johnson J Shiloach R Fass D L Longo S H Bridges 《Journal of immunology (Baltimore, Md. : 1950)》1991,146(6):1806-1811
Treatment of normal mice with a mAb to CD4 (GK1.5) was explored as a means of inhibiting the antibody response to an immunotoxin. Three days of pretreatment with 200 micrograms of GK1.5 completely abrogated the primary antibody response to a 3-micrograms dose of a mutant diphtheria toxin conjugated to an anti-transferrin receptor antibody. The same dose and schedule of anti-CD4 antibody significantly reduced and delayed, but did not prevent, the anamnestic antitoxin response in animals that had been previously primed to the immunotoxin. Three daily injections of anti-CD4 antibodies followed by weekly doses of immunotoxin resulted in a 3-wk delay in the development of antitoxin antibodies, and the kinetics of the antitoxin response correlated with the kinetics of recovery of CD4+ T cells in the spleen and lymph nodes. The antitoxin response to repeated doses of immunotoxin was completely abrogated when anti-CD4 antibodies were given every 2 wk throughout the course of immunotoxin treatment. Thus, transient depletion of Th cells during treatment can block the immune response to an immunotoxin. There was no evidence of tolerance induction with this regimen. 相似文献
25.
Formation of the perinuclear theca in spermatozoa of diverse mammalian species: relationship of the manchette and multiple band polypeptides 总被引:1,自引:0,他引:1
The perinuclear theca is a novel cytoskeletal consisting of a densely layered lamina that surrounds the nucleus of mammalian sperm. Using antibodies specific for the multiple band polypeptides present in the perinuclear theca of bull sperm, we show that a heterogeneous group of immunological related proteins are present in the sperm heads of other mammals with greatly different morphologies, including guinea pig, hamster, rat, and mouse. In none of the species were identical groups of immunoreactive polypeptides found, although immunoreactive proteins of molecular weights 65,000 to 80,000 were present in the sperm heads of all species examined. Immunoreactive proteins less than Mr 55,000 were prominent in rat sperm heads and mouse sperm: guinea pig, hamster, and rat sperm heads and mouse sperm had one band in common at approximately Mr 50,000. Different immunoreactive proteins were present in isolated sperm tails. The perinuclear theca first appeared in the subacrosomal space of round to elongating spermatids. Later, with the caudal movement of the manchette, the postacrosomal segment of the perinuclear theca was deposited in a cephalad to caudal direction along the sperm nucleus. Concomitantly, the cytoplasmic space between the nuclear envelope and the plasma membrane narrowed such that only the theca occupied this portion of the sperm head. Immunoreactivity accompanied the ultrastructural appearance of the subacrosomal layer and the postacrosomal segment. The periods of spermiogenesis, in which sub- and post-acrosomal components of the perinuclear theca are formed and the morphogenesis of sperm organelles with which these elements are associated, suggest that components of this cytoskeletal structure function to join the acrosome and the postacrosomal plasma membrane to the nucleus. 相似文献
26.
Donor-type activated natural killer cells promote marrow engraftment and B cell development during allogeneic bone marrow transplantation. 总被引:5,自引:0,他引:5
W J Murphy M Bennett V Kumar D L Longo 《Journal of immunology (Baltimore, Md. : 1950)》1992,148(9):2953-2960
Purified NK cells were obtained from mice with severe combined immune deficiency and were activated with human IL-2 (hrIL-2) in vitro to determine if, once activated, these cells could be transferred with compatible bone marrow cells (BMC) and promote marrow engraftment in irradiated allogeneic recipients. After culture with hrIL-2, these cells maintained a phenotypic and lytic spectrum consistent with a pure population of activated NK cells. These activated NK cells were then adoptively transferred with the donor BMC and rhIL-2 into lethally irradiated allogeneic hosts. The addition of NK cells with the BMC allowed for more rapid hematopoietic engraftment as determined through short term studies, and greater donor-derived chimerism with accelerated reconstitution of the B cell population as determined with long term analysis. No evidence of graft-vs-host disease was detected in the recipients receiving the activated NK cells with allogeneic T cell replete BMC and hrIL-2. The mechanism by which the transferred NK cells improved BMC engraftment was at least partly through the abrogation of the host effector cell's ability to mediate resistance to the marrow graft. Thus, the administration of donor-type activated NK cells with BMC and hrIL-2 may significantly augment hematopoietic engraftment and immune reconstitution in the clinical setting of allogeneic BMT without giving rise to graft-vs-host disease. 相似文献
27.
C M Loeffler M J Smyth D L Longo W C Kopp L K Harvey H R Tribble J E Tase W J Urba A S Leonard H A Young 《Journal of immunology (Baltimore, Md. : 1950)》1992,149(3):949-956
The causes of the decreased immune responsiveness in tumor-bearing hosts are incompletely understood. The impact of a decreased immune response in cancer patients on the clinical response in immunotherapy trials has not been evaluated. The present report demonstrates a marked decrease in the therapeutic efficacy of adoptively transferred T lymphocytes obtained from murine hosts bearing tumor for greater than 30 days [late tumor-bearing mice (TBM)] as compared with normal mice and mice bearing tumor for less than 21 days (early TBM). In vitro analysis of the functions of the T lymphocytes from late TBM showed an apparently normal proliferative response to anti-CD3 and IL-2 with adequate lymphokine production from CD4+ cells, but a significant decrease in the cytotoxic function of CD8+ cells. The decreased cytotoxicity was not because of cell-mediated suppression. The expression of granzyme B mRNA was significantly delayed and decreased in magnitude in CD8+ cells from late TBM. Culture supernatants from two unrelated tumor cell lines were able to inhibit the cytotoxic activity of normal CD8+ cells in vitro. The tumor-derived suppressive factor is not transforming growth factor-beta (TGF-beta), but it has not been further characterized. The data suggest that one potential mechanism responsible for immunologic defects in patients with large tumor burdens is a tumor-induced defect that compromises the function of CD8+ effector T cells. 相似文献
28.
Calcium accumulation by purified vesicles derived from basolateral membranes of kidney proximal tubules was reversibly inhibited by micromolar concentrations of 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), an inhibitor of anion transport. The inhibitory effect of this compound on Ca2+ uptake cannot be attributed solely to the inhibition of anion transport: (Ca(2+)+Mg2+)ATPase and ATP-dependent Ca2+ transport, respectively. The rate constant of EGTA-induced Ca2+ efflux from preloaded vesicles was not affected by DIDS, indicating that this compound does not increase the permeability of the membrane vesicles to Ca2+. In the presence of DIDS, the effects of the physiological ligands Ca2+, Mg2+, and ATP on (Ca(2+)+Mg2+)ATPase activity were modified. The Ca2+ concentration that inhibited (Ca(2+)+Mg2+)ATPase activity in the low-affinity range decreased from 91 to 40 microM, but DIDS had no effect on the Km for Ca2+ in the high-affinity, stimulatory range. Free Mg2+ activated (Ca(2+)+Mg2+)ATPase activity at a low Ca2+ concentration, and DIDS impaired this stimulation in a noncompetitive fashion. The inhibition by DIDS was eliminated when the free ATP concentration of the medium was raised from 0.3 to 8 mM, possibly due to an increase in the turnover of the enzyme caused by free ATP accelerating the E2----E1 transition, and leading to a decrease in the proportion of E2 forms under steady-state conditions. Alkaline pH totally abolished the inhibition of the (Ca(2+)+Mg2+)ATPase activity by DIDS, with a half-maximal effect at pH 8.3.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
29.
Dirceu de B. Corréa Lourdes F.B. Guerra Otto R. Gottlieb J.Guilherme S. Maia 《Phytochemistry》1981,20(2):305-307
The trunk wood of Qualea labouriauana contains, besides (2R)-5,7,4′-trihydroxy-3′-methoxy-6,8-dimethylflavanone, (2R)-5,7,4′-trihydroxy-8-methylflavanone, the biosynthetically interesting 2,2′-dihydroxy-4,6,4′,6′-tetramethoxy-3,3′-dimethylbenzophenone. From the trunk wood extract of Q. paraensis the first named flavanone crystallized out directly. 相似文献
30.
Surface alterations of fertilized surf clam (Spisula solidissima) eggs induced by concanavalin A 总被引:1,自引:0,他引:1
F J Longo 《Developmental biology》1979,68(2):422-439
Fertilized Spisula eggs, incubated in ConA, were examined at periodic intervals to determine the effects of lectin binding on events of fertilization and cleavage. ConA was localized to specific regions of the vitelline layer and plasma membrane by reacting lectin-treated eggs with horseradish peroxidase and diaminobenzidine. In contrast to eggs, little reaction product was associated with the plasma membrane of spermatozoa. Sperm that fused with ConA-treated eggs failed to move into the cortex of the ovum and were observed as bulbous appendages at the surface of the zygote. Reorganization of sperm nuclei was inhibited, and male pronuclei failed to develop. ConA also inhibited polar body formation and cleavage. The maternally derived chromatin underwent meiosis, and the chromosomes normally taken into the first and second polar bodies were retained within the zygote. All of the maternally derived chromatin was organized within four or more female pronuclei which subsequently entered mitosis. The effects of ConA binding on events at the surface of fertilized Spisula eggs were abrogated by α-methyl-d-mannoside; succinyl-ConA only partially inhibited fertilization-related processes. The effects of ConA are discussed in terms of possible cross-linking of surface components of fertilized Spisula eggs which may inhibit deformation of the zygote cortex. 相似文献