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  • 11
    ISSN: 0006-3592
    Keywords: RGD ; FMDV ; internalization ; integrins ; cell binding ; β-galactosidase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The G-H loop of foot-and-mouth disease virus is a disordered protrusion of the VP1 protein exposed on the virion surface. This short stretch includes an arginine-glycine-aspartic acid tripeptide, a recognized integrin-binding motif, which is responsible for cell attachment and infection. Eight copies of a peptide reproducing the amino acid sequence of this FMDV ligand have been displayed in solvent-exposed regions on an enzymatically active recombinant β-galactosidase. This viral peptide segment enables the chimeric enzyme to bind mammalian cell lines with different efficiencies, probably depending on the number of suitable cell receptors present on each of them. Moreover, it also promotes the internalization of the attached enzyme, which is transiently active inside the cells. These results suggest further exploration of the potential use of short adhesion peptides of viral origin as cell attachment tags to direct the targeted delivery of both genes and enzymes, instead of whole, infectious viruses. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:294-301, 1998.
    Additional Material: 6 Ill.
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  • 12
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Additional Material: 4 Tab.
    Type of Medium: Electronic Resource
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  • 13
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 237 (1993), S. 400-407 
    ISSN: 0003-276X
    Keywords: Human embryo ; Crown-rump length ; Greatest length ; Computer analysis ; Vaginal ultrasound ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Accurate information on the normal growth rate of the human embryo is fundamental to a better understanding of the embryonic period of pregnancy. Crown-rump length measured previously in utero (N = 227) with vaginal ultrasound in 107 in vitro fertilization (IVF) or gamete intrafallopian transfer (GIFT) singleton pregnancies was compared to the greatest length of fixed human embryos from the Carnegie collection, of known developmental stage whose postovulatory ages were estimated from menstrual histories. Average crown-rump length in utero was 60% of the greatest length of the fixed specimens prior to postovulation day 33, but were equal after postovulation day 40. The growth rate of in utero embryos and fixed specimens, analyzed by computer using exponential equations, was compared to linear and polynomial equations used in previously published embryo growth tables. The exponential equation, length = exp(a + b/age), fit in utero measurements best, while the equation length = exp[a + b/exp(age)] fit the fixed specimens best. Differences between length in utero and in fixed specimens may be related to distortion of the fixed embryos resulting from the formalin fixation, to ultrasound distortion, to curling of the embryo, or to incorrectly estimated ages of the fixed specimens. Study of human embryos in utero is now practical with vaginal ultrasound. © 1993 Wiley-Liss, Inc.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
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  • 14
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 210 (1984), S. 491-502 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The ultrastructure of the parotid gland was examined in the little brown bat. The seromucous acinar cells contained abundant granules of variable morphology. These granules were characterized by a submembranous dense layer consisting of fine parallel slats. In some bats, the matrix of the granules was structureless, whereas in others it consisted of closely packed but randomly arranged bundles of tubules. The intercalated ducts had a highly developed rough endoplasmic reticulum, often containing large numbers of intracisternal granules. In contrast, only a few secretory granules were present in the supranuclear cytoplasm. The striated ducts, which exhibited the characteristic basal striations consisting of vertically oriented mitochondria and highly folded plasmalemmas, contained numerous small dense granules in a subluminal band. These granules had a paracrystalline substructure with a periodicity of 8 nm. Excretory ducts strongly resembled striated ducts. They showed the same kind of basal striations and about half their constituent cells contained small paracrystalline granules.
    Additional Material: 16 Ill.
    Type of Medium: Electronic Resource
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  • 15
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The fine structure of corpora lutea from 14 white-tailed deer was studied from early through near-term pregnancy. The corpora contained both thecal and granulosal lutein cells. The small, elongate and cylindrical thecal lutein cells contained many lipid droplets, juxtanuclear Golgi elements, abundant agranular endoplasmic reticulum, and other cytoplasmic organelles. These cells were observed throughout pregnancy. Some of the thecal lutein cells became modified during early to midpregnancy. The modified thecal lutein cells possessed many lipid droplets, several lysosomes, packets of PAS-positive glycogen granules, and numerous small membranous whorls of agranular endoplasmic reticulum. The granulosal lutein cells had highly folded and/or ruffled plasma membranes, abundant tortuous tubular and cisternal agranular endoplasmic reticulum, numerous rod-shaped to round mitochondria with tubular and lamellar cristae, Golgi elements, and a few packets of granular endoplasmic reticulum. Many granulosal lutein cells became modified during mid- to near-term pregnancy by the addition of numerous small membrane-bound osmiophilic droplets and variable numbers of large nonmembrane-bound lipid droplets. The granulosal and thecal lutein cells appeared more active in steroid biosynthesis than the modified granulosal and thecal lutein cells.
    Type of Medium: Electronic Resource
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  • 16
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Serum iron transfer from mother to fetus was measured in near-term pregnant females from 16 species representing 15 families and 7 orders of eutherian mammals. The amount of transfer at 2 hr was 5% or greater in species with hemochorial placentation and less than 0.1% at 3 hr in species with epitheliochorial or endotheliochorial placentation.
    Additional Material: 2 Tab.
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  • 17
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 141 (1989), S. 410-419 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Addition of mitogens to quiescent cells results in rapid ionic changes in the cytoplasm, including pH. We studied the changes in cytoplasmic pH in single Swiss 3T3 cells upon serum stimulation using fluorescence ratio imaging microscopy. Quiescence was attained using two approaches, serum deprivation of subconfluent cells and confluence. All measurements were made in the presence of bicarbonate and the absence of other organic buffers. We also used BCECF coupled to dextran to avoid several artifacts associated with using BCECF-AM, including leakage and phototoxicity. Analysis of the changes in cytoplasmic pH demonstrated a dramatic heterogeneity in the responses of single cells. There were six basic classes of responses, (1) a fast alkalinization, reaching a maximum pH in ∼2-5 min; (2) a slow alkalinization, reaching a maximum pH in 10-20 min; (3) a very slow alkalinization, not reaching a plateau pH within the measurement time; (4) no apparent change in pH during the measurement time; (5) an early transient acidification, followed by either a fast or slow alkalinization; and (6) an acidification, followed by alkalinization and then by a decrease to some intermediate pH. Subconfluent cells exhibited greater heterogeneity in response than confluent cells, with no single dominant class of response. The dominant (55%) response for confluent cells was a gradual alkalinization of ∼0.01 pH units/min. A larger proportion (52%) of subconfluent cells exhibited an early transient acidification compared to confluent cells (7%). A significant proportion of both types of cells (23% subconfluent, 36% confluent) exhibited no change in cytoplasmic pH upon stimulation. In general, the kinetics of changes in cytoplasmic pH were significantly different from the published results with population averaging methods.
    Additional Material: 6 Ill.
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  • 18
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 18 (1982), S. 433-445 
    ISSN: 0730-2312
    Keywords: mitogenicity ; Schwann cells ; axons ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Previous studies in this laboratory have shown that membranes derived from dorsal root ganglia (DRG) neurites are mitogenic for cultured Schwann cells derived from the same source [Salzer et al (1980): J Cell Biol 84:767-778]. Improved procedures are described for preparing Schwann cells derived from dorsal root ganglia that are highly responsive to various mitogens. Under these conditions, the cells respond not only to the neurite mitogen but also to pituitary extracts, dibutyryl cyclic AMP, and cholera toxin that have been shown previously to be good mitogens for Schwannn cells derived from sciatic nerve [Raff et al (1978): Cell 15:813-822], thus reconciling discrepancies in the response of these different Schwann cell preparations to mitogens. Searching for a source of membranes more suitable for biochemical characterization of the neurite mitogen, we found that bovine brain axolemma, prepared by the method of DeVries et al [(1977): Brain Res 147:339-352] is highly mitogenic for Schwann cells. The milotic index of Schwann cells was increased by the addition of axolemma from 0.5%-2% to 30%-50% during 24-h incubation with [3H]thymidine. Half maximal effect was obtained at about 0.4 μg axolemma protein per microwell containing 2-4 × 10 3 cells. The axolemma mitogen appears to be an integral membrane protein that remains bound to the membrane under various ionic conditions but can be extracted in a partially active form with deoxycholate. Like the DRG neurite mitogen, the mitogenic activity of axolemma was abolished by trypsin treatment. Unlike the neurite preparation, however, the mitogenic activity of axolemma was only partially inactivated by heat treatment (60%-70% inactivation). A significant difference between the mitogenic activity of axolemma membranes and neurite membranes is the fact that axolemma membranes fail to stimulate Schwann cell proliferation in a defined, serum-free medium (N-2), whereas neurites show significant mitogenic activity in this medium. These findings indicate a possible difference between DRG neurites and brain axolemma either in the mitogen itself or surface components responsible for recognition between the membranes and the cells.
    Additional Material: 2 Ill.
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  • 19
    ISSN: 0730-2312
    Keywords: protein kinase C ; flow cytometry ; image cytometry ; fluorescence anisotropy ; fluorescence recovery after photobleaching ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Protein kinase C (PKC) has a prominent role in signal transduction of many bioactive substances. We synthesized the fluorescent derivative, phorbol-13-acetate-12-N-methyl-N-4-(N,N′-di(2-hydroxyethyl)amino)-7-nitrobenz-2-oxa-1,3-diazole-aminododecanoate (N-C12-Ac(13)) of 12-O-tetradecanoylphorbol-13-acetate (TPA) to monitor the location of phorbol ester binding sites and evaluate its potential use as a probe of PKC in viable cells. The excitation maximum wavelength of N-C12-Ac(13) is close to 488 nm, facilitating its use in argon-ion laser flow and imaging cytometry. When incubated with 100 nM N-C12-Ac(13) at 25°C, P3HR-1 Burkitt lymphoma cells accumulated the dye rapidly, reaching maximum fluorescence within 25 min, 20-fold above autofluorescence. Addition of unlabeled TPA significantly decreased the fluorescence of N-C12-Ac(13) stained cells in a dose-dependent manner indicating specific displacement of the bound fluoroprobe. Competitive displacement of [3H]-phorbol-12,13-dibutyrate ([3H]-PBu2) from rat brain cytosol with N-C12-Ac(13) gave an apparent dissociation constant (Kd) of 11 nM. N-C12-Ac(13) possessed biological activity similar to TPA. Like TPA (final concentration 65 nM) N-C12-Ac(13), at a lower concentration (51 nM), induced expression of Epstein-Barr viral glycoprotein in P3HR-1 cells, differentiation of promyelocytic HL60 cells, and caused predicted changes in the mitotic cycle of histiocytic DD cells. Microspectrofluorometric images of single cells labeled with N-C12-Ac(13) showed bright fluorescence localized intracellularly and dim fluorescence in the nuclear region, consistent with dye binding mainly to cytoplasmic structures and/or organelles and being mostly excluded from the nucleus. Because of the high level of non-specific binding of N-C12-Ac(13), this probe is not ideal for visualizing PKC in intact cells, but would be a valuable fluoroprobe to investigate the kinetic properties of purified PKC. Also, knowledge gained from these studies allows us to predict structures of fluorescent phorbols likely to have less non-specific binding and, consequently, be potentially useful for monitoring PKC in viable cells.
    Additional Material: 8 Ill.
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  • 20
    ISSN: 0730-2312
    Keywords: NK/A-NK cells ; multicatalytic proteinase complex ; proteasome ; proteases ; enzyme purification ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We have recently described nongranular, cytosolic, high-molecular-weight trypsin-like (A-NKP 1) and chymotrypsin-like (A-NKP 2) proteases of interleukin-2-activated rat natural killer (A-NK) cells. A functional correlation between the inactivation of A-NKP 2 and the inhibition of rat A-NK cell-mediated cytotoxicity was found. Herein we describe the 6,000-fold purification of A-NKP 2 to apparent homogeneity following: isopycnic sucrose gradient fractionation of postnuclear supernatants, molecular sieve chromatography, and heparin-Sepharose® chromatography. We also report the novel finding that A-NKP 2 as well as A-NKP 1, derived from either rat A-NK cells or the rat NK leukemic cell line CRNK-16, are constituents of the multicatalytic proteinase (MCP/proteasome) complexes of these cells. Characteristic biochemical, biophysical, and electron microscopic/ultrastructural similarity to the rat liver proteasome was observed. However, Western blot analysis using polyclonal antibodies to the rat liver proteasome clearly indicated differences in the rat hepatic proteasome and the CRNK-16-derived proteasomal subunits. The identification, characterization, and purification of A-NKP 1 and A-NKP 2, described herein, now allow for further investigation of the potential role of these proteasome components in NK cell function. Moreover, the proteasome of NK and A-NK cells can now be compared and contrasted to the granzymes of lytic granules with respect to their role in cell-mediated cytotoxicity. © 1994 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
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