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PubMed1984

Autoimmune hemolytic anemia by coexisting anti-I and anti-Fl cold agglutinins.

In association with atypical pneumonia, a patient developed acute severe autoimmune hemolytic anemia. Hemoglobin temporarily was only 7.0 g/100 ml, so that the patient needed red blood cell (RBC) transfusion. Hemolysis was found to be caused by high titer cold agglutinins (CA), which occurred transiently during the acute period of the disease. CA of two different specificities, anti-I and anti-Fl, were demonstrated in the patient's serum. Antibodies of the two specificities were clearly separated by absorption/elution experiments using neuraminidase (RDE)-treated RBC. They were distinguished by serologic means: Both anti-I and anti-Fl react more strongly with adult RBC than with newborn and i adult RBC; in contrast to anti-I, anti-Fl does not agglutinate RDE-treated cells. Inhibition experiments showed that I-active substances prepared from papainized RBC exhibited both I and Fl antigenic activity. By RDE-treatment of I-active substances, Fl-activity was markedly reduced, while I-activity was increased.

Adult↗
PubMed2005

Localization of the Drosophila protein FL(2)D in somatic cells and female gonads.

The Drosophila gene female-lethal(2)d [fl(2)d] has been implicated in the alternative splicing regulation of genes involved in sexual determination, such as Sex-lethal and transformer, and in the alternative splicing of the gene Ultrabithorax. Here, the expression and subcellular localization of the FL(2)D protein is reported. FL(2)D is ubiquitously expressed in embryos, in imaginal discs of larvae and in ovaries. In Schneider cells, the protein is detected throughout the nucleoplasm, excluding the nucleolus, and partially co-localizes with SXL in nuclear speckles. In addition, FL(2)D associates with multiple transcriptionally active loci on salivary gland polytene chromosomes, consistent with the proposal that the protein is involved in the post-transcriptional regulation of a variety of genes. Interestingly, the localization of FL(2)D in nurse cells changes during oogenesis, from a rather diffuse early nuclear pattern in the germarium to a preferential accumulation at the nuclear periphery of nurse cells of developing cysts.

Animals↗
PubMed2000

Probing the cathepsin D using a BODIPY FL-pepstatin A: applications in fluorescence polarization and microscopy.

Redistribution of cathepsin D, a major lysosomal aspartic endopeptidase, has been related to various pathological progressions during tumor formation and oxidation stress. We have synthesized a fluorescent probe for cathepsin D, where the pepstatin A was covalently conjugated with the BODIPY (Boron dipyrromethene difluoride) fluorophore. In vitro, BODIPY FL-pepstatin A inhibits cathepsin D activity with an IC50 of 10 nM. The nature of its binding to cathepsin D was further characterized using a fluorescence polarization measurement. Results showed that BODIPY FL-pepstatin A selectively binds to cathepsin D at pH 4.5. In fixed cells, BODIPY FL-pepstatin A stained lysosomes, where it co-localized with cathepsin D. This staining was depleted when cells were co-incubated with unlabeled pepstatin A in acidic buffer. In live cells, BODIPY FL-pepstatin A is internalized and transported to lysosomes. The staining in the lysosomes can be competed with unlabeled pepstatin A. These properties, along with the good photostability of the BODIPY FL fluorophore, make this probe a novel tool for the study of the secretion and trafficking of cathepsin D.

Animals↗
PubMed1997

Neutralization of the human immunodeficiency virus type 1 primary isolate JR-FL by human monoclonal antibodies correlates with antibody binding to the oligomeric form of the envelope glycoprotein complex.

To test whether antibodies that are neutralizing or nonneutralizing for human immunodeficiency virus type 1 (HIV-1) primary isolates can be distinguished by their affinities for the oligomeric envelope glycoproteins, we selected HIV-1(JR-FL) as a model primary virus and a panel of 13 human monoclonal antibodies (MAbs) and evaluated three parameters: (i) half-maximal binding to recombinant monomeric envelope, gp120(JR-FL); (ii) half-maximal binding to oligomeric envelope of HIV-1(JR-FL) expressed on the surface of transfected 293 cells; and (iii) neutralization of HIV-1(JR-FL) in a peripheral blood mononuclear cell-based neutralization assay. Two conclusions can be drawn from these experiments. First, we confirm that antibody interactions with monomeric gp120 do not predict primary virus neutralization. Second, we show that neutralization correlates qualitatively with the relative affinity of an antibody for the oligomeric envelope glycoproteins, at least for HIV-1(JR-FL).

Antibodies, Monoclonal↗
PubMed1991

A hybrid form of alkaline phosphatase produced in FL-amnion cells.

FL-amnion cells have mainly two alkaline phosphatase (AP) isozymes, of which the faster migrating one (FL-APF) on 5% polyacrylamide gel electrophoresis has proved to be identical to the Kasahara isozyme, a tumor-associated AP of intestinal type. But the other slower migrating one (FL-APs) remains to be characterized. Immunological and enzymic examination of purified FL-APs revealed that it is a hybrid form of AP consisting of one subunit with a molecular weight of 66,200 coming from the same subunit as that of FL-APF and another one with a molecular weight of 58,000 from placental AP subunit with modified glycosylation.

Alkaline Phosphatase↗
PubMed2003

[Ex vivo expansion of CD34+ cells from human umbilical cord blood by FL gene transfected bone marrow stromal cells].

The object of this study was to explore the effect of FL gene transfected marrow stromal cells on expansion of human cord blood CD34(+) cells ex vivo. A co-culture system was established with FL gene transfected human marrow stromal cells and cord blood CD34(+) cells. The CD34(+) cells were cultured in different culture systems with different combinations of stromal cells and SCF, IL-3 and FL. The numbers of total nucleated cells, CFCs and CD34(+) cells were repeatedly counted in culture systems for 3 weeks. The results showed that the FL transgenic marrow stromal cell feeder layer significantly enhanced the expansion of total nucleated cells, CFC and CD34(+) cells, compared with stromal cell feeder layer and stromal cell-free culture systems. The transgenic stromal cells and cytokines co-culture system manifested the most potent expansion of the cord blood cells with increase of the number of total nucleated cells by (122.5 +/- 4.3)-fold, CFCs by (39.6 +/- 2.7)-fold and CD34(+) cells by (11.8 +/- 0.52)-fold. It is concluded that human umbilical cord blood CD34(+) cells can be extensively expanded ex vivo by using FL gene transfected stromal cells combined with cytokines.

Antigens, CD34↗
PubMed1987

Serological relationship between mouse adenovirus strains FL and K87.

Three-week-old outbred mice were inoculated intraperitoneally or orally and intranasally with the FL or K87 strains of mouse adenovirus and bled at intervals after infection. Serum was tested by both the complement fixation and indirect immunofluorescence tests for reactivity with either virus antigen. A unilateral relationship was detected between FL and K87 strains. Serum from mice given the FL strain of virus reacted in both tests with FL and K87 antigens. Serum from mice given the K87 strain reacted only with the homologous antigen. Serum antibody titers were generally higher in the immunofluorescence test than in the complement fixation test. These observations stress the need to use both FL and K87 antigens for specific serologic diagnosis of adenovirus infection in mouse colonies.

Adenoviridae↗
PubMed1992

Studies of composite grafts of fetal pancreas (FP) and fetal liver (FL) in the streptozotocin-induced diabetic rat.

A hepatotrophic effect of pancreatic islets on co-transplanted hepatocytes has been described recently by Ricordi et al. We investigated a possible reciprocal effect of co-transplanted fetal liver (FL) on fetal pancreas (FP) isografted into streptozotocin diabetic rats. FL was co-transplanted with FP in three sites: the new intramural small bowel (ISB) site, the conventional renal subcapsular (RSC) site, and the historically inhospitable intramuscular (IM) site. Overall, as compared to grafts of FP alone, composite FP/FL grafts consistently provided earlier restoration of normoglycemia in streptozotocin diabetic recipients (24 +/- 8 vs. 67 +/- 43 days P = 0.001). The proportion of recipients rendered normoglycemic and the clearance of glucose was site-dependent. For FP and FP/FL recipients respectively, the ISB site resulted in 100% normoglycemia in both groups (19/19 and 6/6), the RSC site resulted in 71% (5/7) and 40% (2/5) and the IM site resulted in 14% (1/7) and 67% (6/9). In normoglycemic recipients, glucose clearance was normal or supraphysiologic except for the RSC site. Composite isografts brought about normoglycemia in 75% (6/8) of recipients treated with beta-cell toxic doses of cyclosporine that prevented reversal of diabetes in recipients of FP alone. Co-transplantation of FL benefits FP through paracrine mechanisms mediated by unknown factor(s). Thus, as compared to grafts of FP alone, composite FP/FL grafts established normoglycemia more rapidly, mitigated cyclosporine toxicity and corrected diabetes when transplanted in the small bowel site. There are several mediators that may be responsible for these paracrine effects between the liver cells and the pancreas. IGF-1 elaborated by FL is the most likely trophic factor.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Comparative biological characterization of mouse adenovirus strains FL and K 87 and seroprevalence in laboratory rodents.

The growth, stability and seroprevalence in laboratory rodents of the two known strains of mouse adenovirus were compared. The FL strain of mouse adenovirus grew in both L 929 murine fibroblasts and in CMT-93 murine rectal carcinoma cells, whereas the K 87 strain grew only in CMT-93 cells. The bulk of the FL progeny virus was released from the host cells. K 87 virus was largely cell-associated. Both virus strains were stable at 37 degrees C in liquid medium. The K 87 strain was completely inactivated after 5-15 minutes at 56 degrees C, whereas FL infectivity was still detected after two hours at this temperature. Both virus strains were stable in the dessicated state for 14 days, although FL viability was more dependent on the presence of protein in the virus diluent. Seroepidemiologic data suggest that viruses antigenically related to mouse adenovirus are more prevalent among laboratory rats than among laboratory mice and that the virus(es) infecting rats differ from those infecting mice. Results of retrospective serologic testing suggest an association between mouse adenovirus and an outbreak of disease in a mouse breeding colony.

Adenoviridae↗
PubMed1986

Binding of cardiotoxin analogue III from Formosan cobra venom to FL cells.

The binding equilibrium at 37 or 0 degrees C of 125I-cardiotoxin analogue III (CT III) to fetal lung (FL) cells (cultured human amnion cells) was achieved within 1 h, and the binding at 37 degrees C was irreversible. The Scatchard analysis at 37 degrees C on the binding of 125I-CT III indicated that FL cells had two types of binding sites with different association constants. The association constant and the number of high-affinity sites was 1.1 X 10(10) mol-1 or 2.8 X 10(6) per FL cell, respectively. At 37 or 0 degrees C, the cytotoxicity of CT III paralleled the amount of bound CT III to FL cells, and at 37 degrees C was inhibited by the presence of acidic phospholipids.

Binding Sites↗
PubMed1984

Appearance of cytotoxic antibody to viral gp70 on feline lymphoma cells (FL-74) in cats during ex vivo immunoadsorption therapy: quantitation, characterization, and association with remission of disease and disappearance of viremia.

Fifty cats with feline leukemia virus (FeLV) infection and leukemia-lymphoma complex were treated by ex vivo immunoadsorption with Staphylococcus protein A-bound filters. Most cats responded to therapy. Twelve showed tumor regression, including disappearance of tumor cells, but died later of other complications. Three have had long-term remission of more than 1 year and remain healthy. A consistent finding in these three cats was the appearance during treatment of a complement-dependent cytotoxic antibody against cat lymphoma cells (FL-74). The cytotoxic antibody increased substantially during treatment. Appearance and increase of the cytotoxic antibody was associated with disappearance of FeLV from blood and remission of leukemia. By electroblot analysis, antibody to FeLV protein (Mr, 70,000) was detected in serum prior to detection of the cytotoxic antibody. The cytotoxic antibody was found by immunofluorescence to be specific for antigens on membranes of viable FL-74 cells. By using monoclonal antibodies to FL-74 cells and to components of FeLV, the cytotoxic antibody was shown to be directed against gp70, a glycoprotein of Mr 70,000, but not against p27 of FeLV or other membrane antigen(s) of FL-74 cells. The development of a high concentration of cytotoxic antibody to FeLV gp70 may play an important role in tumor regression and in disappearance of FeLV infection.

Animals↗
PubMed1975

Attachment of lipoprotein to murein (peptidoglycan) of Escherichia coli in the presence and absence of penicillin FL 1060.

In vivo studies on the attachment of lipoprotein to the murein (peptidoglycan) of Escherichia coli showed that it takes several generations of growth until the amount of lipoprotein on newly made murein is equilibrated. The technique used involves degradation of the sodium dodecyl sulfate-insoluble murein-lipoprotein complex (sacculus, rigid layer) with lysozyme and separation of the labeled products on paper. No lipoprotein was found on murein subunits incorporated during a pulse of [3H]diaminopimelate for 1 min in logarithmically growing cells at 37 C. Even after one doubling of the cell mass, only 4 to 8% of the labeled murein was isolated as bound to lipoprotein. With uniformly labeled murein, 30% remains bound to lipoprotein after lysozyme treatment, corresponding to three murein subunits. Therefore it can be concluded that during pulse labeling either no lipoprotein is incorporated into the newly synthesized murein or no murein subunits are inserted into existing murein around lipoprotein attachment sites. Longer pulse and pulse-chase experiments argue for the latter interpretation. It is therefore concluded that incorporation of murein subunits into the growing murein polymer is not at all a random process. Instead, quite large areas of murein, on which lipoprotein is situated, seem to be preserved. Under the influence of penicillin FL 1060 murein synthesis is 50% inhibited. The rate of lipoprotein attachment is less affected so that increasing amounts of lipoprotein become attached during spheroplast formation. By the time the stationary growth phase has been reached, the lipoprotein content of the murein has doubled. Diaminopimelate auxotrophic mutants require, in the presence of penicillin FL 1060, more diaminopimelate for full growth than in the absence of penicillin FL 1060. This finding and the fact that murein synthesis is always inhibited by 50% over a wide range of penicillin concentration (1 to 1,000 mug/ml) point to the inhibition of an enzymatic step of murein synthesis which can be partially bypassed by a second enzyme, less efficient but resistant to penicillin FL 1060.

Binding Sites↗
PubMed1993

Protection of cats against feline leukemia virus by vaccination with a canarypox virus recombinant, ALVAC-FL.

Two ALVAC (canarypox virus)-based recombinant viruses expressing the feline leukemia virus (FeLV) subgroup A env and gag genes were assessed for their protective efficacy in cats. Both recombinant viruses contained the entire gag gene. ALVAC-FL also expressed the entire envelope glycoprotein, while ALVAC-FL(dl IS) expressed an env-specific gene product deleted of the putative immunosuppressive region. Although only 50% of the cats vaccinated with ALVAC-FL(dl IS) were protected against persistent viremia after oronasal exposure to a homologous FeLV isolate, all cats administered ALVAC-FL resisted the challenge exposure. Significantly, protection was afforded in the absence of detectable FeLV-neutralizing antibodies. These results represent the first effective vaccination of cats against FeLV with a poxvirus-based recombinant vector and have implications that are relevant not only to FeLV vaccine development but also to developing vaccines against other retroviruses, including human immunodeficiency virus.

Animals↗
PubMed1974

Activity of FL 1060, a new beta-lactam antibiotic, against urinary tract pathogens.

The effect of the new beta-lactam antibiotic FL 1060 on urinary tract pathogens, of which 71 were sensitive to and 100 resistant to ampicillin, is presented here. When tested by conventional methods FL 1060 was found to be highly active against ampicillin-sensitive strains of E. coli but phenotypically resistant variants readily emerged, particularly when a large inoculum was used. Thirty-one per cent of ampicillin-resistant strains were found to be sensitive to less than 500 mug FL 1060 per ml, and in general this antibiotic was more active than ampicillin against resistant strains. In a system simulating the mechanical features of the urinary bladder the activity of FL 1060 against three sensitive strains of E. coli was found to be greater than ampicillin in conditions of low, but not high, osmolality.

Ampicillin↗
PubMed2002

The fluorescent probe Bodipy-FL-verapamil is a substrate for both P-glycoprotein and multidrug resistance-related protein (MRP)-1.

Several fluorescent probes have been used in functional studies to analyze drug transport in multidrug-resistant cells by fluorescent microscopy. Because many of these molecules have some drawbacks, such as toxicity, nonspecific background, or accumulation in mitochondria, new fluorescent compounds have been proposed as more useful tools. Among these substances, Bodipy-FL-Verapamil, a fluorescent conjugate of the drug efflux blocker verapamil, has been used to study P-glycoprotein activity in different cell types. In this study we tested by fluorescent microscopy the accumulation of Bodipy-FL-Verapamil in cell lines that overexpress either P-glycoprotein (P-gp) or multidrug resistance-related protein 1 (MRP1). Expression of P-gp and MRP1 was evaluated at the mRNA level by RT-PCR technique and at the protein level by flow cytometric analysis using C219 and MRP-m6 monoclonal antibodies. Results indicate that Bodipy-FL-Verapamil is actually a substrate for both proteins. As a consequence, any conclusion about P-gp activity obtained by the use of Bodipy-FL-Verapamil as fluorescent tracer should be interpreted with caution.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMedDate not supplied

Effects of FL-386 on faecal lipid excretion in humans.

A newly synthesized inhibitor of pancreatic lipase and micelle formation, FL-386, was administered at a dose of 400 mg (in the diet, for seven consecutive days) to nine healthy adult volunteers, and changes in faecal mass, frequency of defaecation, and properties of the stools were observed. High performance liquid chromatography and gas-liquid chromatography were used to analyse the faeces for short-chain carboxylic acids, neutral sterols, bile acids, fats and hydroxyfatty acids. FL-386 had little effect on the amounts and composition of short-chain carboxylic acids, neutral sterols, and bile acids excreted, nor did it produce changes in the composition of fatty acids, or the percentages of hydroxyfatty acids in the stool. However, in those patients treated with FL-386, the faecal mass was increased, and stools were softer and contained increased amounts of fatty acids. The compound did not produce particularly fatty stools. It was concluded that FL-386 induces slight disturbance in the digestion and absorption of dietary lipids.

Adult↗
PubMed1986

[Effects of 3-methyl-4-oxo-2-phenyl-N-[2-piperidinyl) ethyl]-4H-1-benzopyran-8-carboxyamide hydrochloride monohydrate (FL-155), a new anti-pollakiuria agent, on rhythmic bladder contractions in anesthetized rats].

The effects of FL-155, which was synthesized to develop a new orally-active anti-pollakiuria agent, on the rhythmic bladder contractions were studied in anesthetized rats. At a pressure exceeding 10 cm H2O in the bladder, a rhythmic bladder contraction was observed up to at least 120 min. This response was abolished by a spinal (C1 level) cut, cuts of both pelvic nerves, thiopental (3.0 mg/kg, i.v.) or lidocaine (1.0 mg/kg, i.v.); and atropine (0.01 mg/kg, i.v.) strongly inhibited the amplitude of the response. FL-155 and flavoxate, in intravenous (0.3-3.0 mg/kg and 1.0-3.0 mg/kg, respectively) and intraduodenal (12.5-100 mg/kg and 200-400 mg/kg, respectively) administrations, dose-dependently abolished the rhythmic bladder contractions, and FL-155 was 8-16 times more potent than flavoxate in intraduodenal administrations. These results suggest that the rhythmic bladder contraction in anesthetized rat may be a polysynaptic reflex through pelvic nerves and the central nervous system (supraspinal level), and FL-155 appears to be a candidate for an orally active anti-pollakiuria agent.

Animals↗
PubMed2002

Long-term locomotor training up-regulates TrkB(FL) receptor-like proteins, brain-derived neurotrophic factor, and neurotrophin 4 with different topographies of expression in oligodendroglia and neurons in the spinal cord.

Neurotrophins are potent regulators of neuronal survival, maintenance, and synaptic strength. In particular, brain-derived neurotrophic factor (BDNF), acting through full-length TrkB receptor (TrkB(FL)), is implicated in the stimulation of neurotransmission. Physical activity has been reported to increase BDNF expression in the brain and spinal cord. In this study we have evaluated the hypothesis that activation of a spinal neuronal network, due to exercise, affects the entire spinal neurotrophin system acting via TrkB receptors by modulation of BDNF, neurotrophin 4 (NT-4), and their TrkB receptor proteins. We investigated the effect of treadmill walking (4 weeks, 1 km daily) on distribution patterns and response intensity of these proteins in the lumbar spinal cord of adult rats. Training enhanced immunoreactivity (IR) of both neurotrophins. BDNF IR increased in cell processes of spinal gray matter, mainly in dendrites. NT-4 IR was augmented in the white matter fibers, which were, in part, of astrocytic identity. Training strongly increased both staining intensity and number of TrkB(FL)-like IR small cells of the spinal gray matter. The majority of these small cells were oligodendrocytes, representing both their precursor and their mature forms. In contrast, training did not exert an effect on expression of the truncated form of TrkB receptor in the spinal cord. These results show that both neuronal and nonneuronal cells may be actively recruited to BDNF/NT-4/TrkB(FL) neurotrophin signaling which can be up-regulated by training. Oligodendrocytes of the spinal gray matter were particularly responsive to exercise, pointing to their involvement in activity-driven cross talk between neurons and glia.

Animals↗