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J M Shields

Publications and source records attributed to J M Shields.

18 recordsLinked to original sources
PubMed1994

Galactose consumption, metabolism, and follicle-stimulating hormone concentrations in women of late reproductive age.

OBJECTIVE: To test the hypothesis that high galactose consumption and low activity of galactose-1-phosphate uridyl transferase (transferase) is associated with early ovarian senescence among nongalactosemic women. DESIGN: Cross-sectional study. Data collection consisted of a self-administered questionnaire with sections on diet (food frequency data to measure galactose consumption), reproductive, and medical histories. One blood sample was collected to measure FSH and transferase activity; FSH was used as a measure of ovarian senescence. Among women who were having menstrual periods at least every 8 weeks, the blood sample was drawn in the early follicular phase (days 2 to 4) of a menstrual cycle. PARTICIPANTS: Two hundred ninety-five women volunteers ages 38 to 49 years who had not had a hysterectomy or oophorectomy were recruited through posters and advertisements. MAIN OUTCOME MEASURE: Serum FSH concentrations. RESULTS: Controlling for age, smoking, and body mass, transferase activity and FSH were unrelated. However, FSH levels were 29% higher (95% confidence intervals, 9% to 52%) among women who reported consuming > or = 6 g galactose/d. CONCLUSION: These data do not support the hypothesis that low transferase activity represents a genetic predisposition for early ovarian senescence, as measured by FSH levels in women ages 38 to 49 years. However, the hypothesized positive association between galactose consumption and FSH was supported.

Adult↗
PubMed1992

Collision between law and ethics: consent for treatment with adolescents.

The dilemmas between legal obligations and ethical responsibilities can often create problems in clinical work. The treatment of minors, and particularly adolescents, can present special issues to the clinician that are becoming increasingly frequent and difficult. The issue of informed consent for treatment of adolescents raises serious questions for the clinical practitioner who is faced with both legal and ethical dilemmas in making decisions about treatment. There are an increasing number of cases where adolescents may seek treatment yet are in circumstances that preclude parental consent. This paper uses case material to illustrate some of the legal, ethical, and treatment considerations in the situation of adolescent treatment where parental consent is problematic.

Adolescent↗
PubMed1990

High-resolution immunogold localization of Giardia cyst wall antigens using field emission SEM with secondary and backscatter electron imaging.

We describe here the ultrastructural localization of Giardia cyst antigens in the filaments associated with the outer portion of intact cysts and on developing cyst wall filaments in encysting trophozoites. Post-embedding immunogold labeling of thin sections of intact Giardia cysts with polyclonal and monoclonal antibodies specific for cyst wall antigens (major protein bands of approximately 29, 75, 88, and 102 KD on Western blots) showed strong labeling of the filamentous cyst wall, whereas no labeling was seen on the membranous portion. High-resolution field emission scanning electron microscopy (FESEM) of Giardia cysts revealed that the cyst wall-specific polyclonal rabbit antisera and monoclonal mouse antibody produced gold labeling of 20-nm filaments in the cyst wall as detected with secondary electron imaging (SEI) and backscatter electron imaging (BEI) at 10 kV, despite coating of the cells with platinum by ion sputtering. FESEM studies of encysting Giardia trophozoites demonstrated that immunostaining with antibodies to cyst wall antigens produced colloidal gold labeling of developing cyst wall filaments on the cell surface; however, the intervening membrane domains were unlabeled. Substitution of normal serum for cyst wall-specific antibodies, or preabsorption of specific antibodies with Giardia cysts, eliminated immunolabeling of the filaments.

Animals↗
PubMed1986

Signal transduction in human neutrophil leucocytes: effects of external Na+ and Ca2+ on cell polarity.

Stimulation of neutrophil leucocytes with chemotactic factors is known to result in membrane permeability changes, as evidenced by fluxes of Na+ and K+ across the cell membrane together with an increased uptake of Ca2+ from the medium. These fluxes have been implicated in the transduction mechanisms of various responses, including locomotion and subsequent chemotaxis. We have previously reported that exposure of unstimulated, round neutrophils held in suspension, to the chemotactic peptide fMet-Leu-Phe confers morphological polarity on the neutrophils by stimulating waves of contraction, which are also intimately connected with locomotion on an appropriate substratum. As the acquisition of polarity is the important first step in the chemotactic response we have investigated the effects of modifying the external ionic environment and of various ion channel blockers on the polarizing response of neutrophils held in suspension. Removal and chelation of both Ca2+ and Mg2+ from the external medium did not inhibit the acquisition of polarity and a variety of inorganic Ca2+ channel blockers together with the organic Ca2+ antagonists, verapamil and D600, were ineffective in inhibiting the response. Replacement of Na+ in the external medium with choline inhibited the polarizing response completely but tetrodotoxin, which blocks fast Na+ channels, and amiloride, which inhibits Na+/K+ exchange, had no effect. Inhibition of the Na+/K+-ATPase with ouabain and also tetraethylammonium ions, which block potassium channels, had no inhibiting effect on polarization. These results indicate that while Ca2+ and Mg2+ are not required in the external medium, Na+ is essential, and therefore Na+/K+ fluxes across the cell membrane play a role in initiating locomotion.

Chemotaxis↗
PubMed1985

Neutrophil leucocyte chemotaxis: a simplified assay for measuring polarizing responses to chemotactic factors.

A method is described which greatly simplifies the screening of compounds which are potentially chemotactic for neutrophil leucocytes. Neutrophils isolated from blood by a standardised procedure are greater than 95% spherical in morphology. Addition of chemotactic factors in isotropic, non-gradient, concentrations induces the spherical shape to become polarized. The degree of polarity depends on the concentration of the factor used, as does the percentage of cells which become polarised. All compounds which induce a good response in assays measuring cell accumulation or orientation in gradients induced a consistent polarizing response in non-gradient conditions with the cells held in suspension. The advantage of this simplified assay over methods currently in use are discussed.

Chemotactic Factors↗
PubMed1985

Behaviour of neutrophil leucocytes in uniform concentrations of chemotactic factors: contraction waves, cell polarity and persistence.

The essential component of any hypothesis of random or directed cell movement is the mechanism of cell polarity. In this paper we describe the polar behaviour of human neutrophil leucocytes in uniform concentrations of chemotactic factors both in suspension and while moving across surfaces. Neutrophils exposed to uniform concentrations of chemotactic factors in suspension around the dissociation constant (Kd) for the receptor rapidly become distinctly bipolar; neutrophils exposed to supraoptimal uniform concentrations (100-fold greater than Kd) of chemotactic factors in suspension, although morphologically active, never reached the same degree of polarity as cells in optimal concentrations. These differences in polarity were shown to be the direct result of equatorial contraction waves stimulated on the cell surface by interaction with chemotactic factors. In optimal concentrations of chemotactic factors, contraction waves were initiated from one region of the cell, whereas in supraoptimal concentrations of chemotactic factors contraction waves emanated from all areas of the cell surface. Asymmetry in the distribution of surface receptors for Fc and C3b were observed in neutrophils polarized in uniform concentrations of chemotactic factor. In neutrophils, motile but not well polarized (in 10(-6) M-N-formylmethionyl-leucyl-phenylalanine (fMLP), receptors were uniformly distributed. In neutrophils polarized in concentrations of fMLP near the Kd for the receptor (10(-8) M) receptors for C3b and Fc were localized in the anterior region of the moving cell. The link between contraction waves, cell polarity and receptor redistribution and their initiation by chemotactic peptides is discussed in the context of neutrophil locomotion and response to chemical signals.

Cell Movement↗
PubMed1984

Contraction waves in lymphocyte locomotion.

In this paper we propose that the constriction ring, a prominent feature of moving leucocytes, is a major source of locomotive force. Analysis of time-lapse films of lymphocytes in suspension and moving through three-dimensional collagen gels, demonstrated that the constriction ring was the morphological manifestation of a wave of circular contraction that moved antero-posteriorly. In lymphocytes in suspension the wave moved, although the cells could not. Analysis of lymphocytes moving through a collagen gel revealed that the waves remained stationary with respect to the external environment while the cell appeared to move forward through them. Passage of a single equatorial contraction wave resulted in cell lengthening: a shortening of the region posterior to the constriction was observed in cells moving through collagen gels, but not in lymphocytes held in suspension, suggesting that attachment of cells to the collagen network was necessary for longitudinal contraction. Lymphocyte attachment to collagen gels was mediated through the rapid extension of bleb-like structures into the collagen network. Transmission electron microscopy (TEM) failed to demonstrate any organized structure at the constriction ring. NBD-Phallacidin staining of lymphocytes together with TEM demonstrated that F-actin was distributed evenly throughout the length of the cell. Cell polarity was clearly recognizable by the distribution of coated vesicles, microvilli, and all organelles to the rear, and Thy 1-2 to the front, of motile cells, but polarity could be reversed by the passage of a single contraction wave starting at the rear of the cell, without prior redistribution of these structures.

Actins↗
PubMed1984

Invasion of collagen gels by mouse lymphoid cells.

Small mouse lymphocytes from lymph nodes rapidly invaded three-dimensional collagen gels (in the absence of any added chemical attractant). In short-term assays (2-8 hr) this property was restricted to 20-25% of the cell population. Invasion was an active process involving cell locomotion. Time-lapse cinematography revealed that movement was erratic with frequent changes in cell speed. Tracks of cell paths within collagen gels demonstrated that lymphocytes made narrow angles of turn and thus showed a 'persistent random-walk' similar to other cell types moving on plane substrata. Analysis of lymphocyte movement within aligned collagen gels demonstrated that locomotion was biased in the axis of fibre alignment, i.e. lymphocytes showed contact guidance. Separated B lymphocytes invaded collagen gels at a slower rate than unseparated lymph node cells, as also did T cells purified by filtration through nylon wool columns. This latter anomaly implied that nylon wool filtration selectively depleted cells with invasive characteristics from a heterogeneous lymphocyte population. A comparison of Peyer's patch and lymph node lymphocytes showed that both populations invaded at the same rate but the latter cell type did this in greater numbers. This difference may reflect the different proportions of B and T lymphocytes in the two tissues. Lymphocytes from oxazolone-stimulated lymph nodes showed greatly increased movement into collagen matrices compared to unstimulated control lymph node lymphocytes. This increase was demonstrated to be a property of the blast cell population by separating the cells on Percoll gradients into lymphoblast-enriched and -depleted populations.

Animals↗
PubMed1983

Separation of mouse lymphoblasts by discontinuous density centrifugation on Percoll gradients.

Mouse lymphoblasts generated in vivo by a topical application of the contact sensitizer oxazolone or by the contents of the gut lumen were separated by discontinuous density centrifugation on Percoll gradients. A 3-step gradient was used to divide the cells into two subpopulations. For cells from oxazolone stimulated lymph nodes, the low density band contained 20-30% of the initial cell number applied to the gradient; 25-40% of this population were in S phase and nearly all the large and pyroninophilic cells were confined to this layer. The high density step cells (70-80% of initial cell number) were predominantly small lymphocytes with less than 0.5% in S phase. Similar results were obtained using cells from picryl chloride stimulated lymph nodes or from mesenteric lymph nodes.

Animals↗
PubMed1983

The orientation of fibroblasts and neutrophils on elastic substrata.

The reaction of fibroblasts to the elastic properties of the substratum was studied using elastic collagen films. These films were stretched in one axis to give a substratum which was anisotropic in its elasticity and deformability. Analysis of the orientation of fibroblasts cultured on these substrata showed that they oriented along the axis of stretch which was also the axis of fibre alignment. This orientation was significantly reduced when the films were made less elastic by attachment to a glass slide and chemical fixation. Neither of these procedures appeared to alter the surface shape of these films, which suggests that the elastic properties of the substratum markedly influence the orientation of fibroblasts. The orientation of locomotion of neutrophil leukocytes on elastic collagen films was also analysed and no bias along the axis of stretch was observed. This was compared with neutrophil locomotion in 3-D stretched collagen gels, in which a strong bias along the axis of stretch and of fibre alignment was observed. The possible reasons for the response of these two cell types is discussed.

Animals↗
PubMed1982

Lymphocyte locomotion and attachment on two-dimensional surfaces and in three-dimensional matrices.

The adhesion and locomotion of mouse peripheral lymph node lymphocytes on 2-D protein- coated substrata and in 3-D matrices were compared. Lymphocytes did not adhere to, or migrate on, 2-D substrata suck as serum- or fibronectin-coated glass. They did attach to and migrate in hydrated 3-D collagen lattices. When the collagen was dehydrated to form a 2-D surface, lymphocyte attachment to it was reduced. We propose that lymphocytes, which are poorly adhesive, are able to attach to and migrate in 3-D matrices by a nonadhesive mechanism such as the extension and expansion of pseudopodia through gaps in the matrix, which could provide purchase for movement in the absence of discrete intermolecular adhesions. This was supported by studies using serum-coated micropore filters, since lymphocytes attached to and migrated into filters with pore sizes large enough (3 or 8 mum) to allow pseudopod penetration but did not attach to filters made of an identical material (cellulose esters) but of narrow pore size (0.22 or 0.45 mum). Cinematographic studies of lymphocyte locomotion in collagen gels were also consistent with the above hypothesis, since lymphocytes showed a more variable morphology than is typically seen on plane surfaces, with formation of many small pseudopodia expanded to give a marked constriction between the cell and the pseudopod. These extensions often remained fixed with respect to the environment as the lymphocyte moved away from or past them. This suggests that the pseudopodia were inserted into gaps in the gel matrix and acted as anchorage points for locomotion.

Animals↗
PubMed1982

Some determinants of the locomotory behaviour of phagocytes and lymphocytes in vitro.

In this review, we discuss some physical and chemical determinants of locomotor behaviour of phagocytic cells and lymphocytes as studied in visual assays, paying particular attention to the following points. (a) A distinction is made between chemokinesis and other forms of kinesis. We propose that the term chemokinesis be reserved for responses resulting from selective recognition of chemical substances. Many kineses in leucocytes may not involve such recognition, but may result from a variety of physical factors, such as those that alter the adhesiveness between cell and substratum. (b) Neutrophils moving in aligned gels of collagen or fibrin show contact guidance of locomotion, i.e. bidirectional movement along the axis of alignment of the fibres of the gel. Thus neutrophils show directional locomotion not only in chemotactic gradients but also in response to physical properties of the substratum. (c) Lymphocytes adhere poorly and move poorly on 2D protein coated substrata. However they move rapidly through 3D collagen gels. This locomotion may be independent of adhesion since lymphocytes may gain traction for locomotion by expanding pseudopods into gaps in the gel matrix, then using the pseudopod as an anchor for subsequent locomotion in any direction.

Animals↗
PubMed1979

Relations between Fc receptor function and locomotion in human lymphocytes.

The relationship between the surface binding sites on human lymphocytes for chemotactic factors and for the Fc fraction of IgG was investigated using both blood lymphocytes and established cultures of human lymphoblasts. Pretreatment of human blood lymphocytes with a variety of chemotactic factors inhibited Fc-rosette formation. This was true even of small formylated peptides, for example, formyl-methionyl-phenylalanine (chemotactic) inhibited Fc-rosetting but unformylated methionyl-phenylalanine (non-chemotactic) did not. Conversely pretreatment of lymphocytes with IgG inhibited their locomotor reactions to a variety of chemoattractants. Aggregated IgG was more inhibitory than non-aggregated IgG and the inhibition was mediated via the Fc piece. In a filter assay, native IgG was chemokinetic but not chemotactic for lymphocytes. Heat-aggregated IgG induced more locomotion of lymphocytes than native IgG, and was possibly chemotactic, but no unequivocally so. The possibility that chemotactic factors and the Fc portion of IgG compete for the same cell surface receptor was investigated by binding studies using cultured lymphoblasts. These studies suggested that the reciprocal inhibition could not be explained by competition for receptors. An alternative explanation was suggested by the finding that inhibition of locomotion by aggregated IgG was dependent on the presence of divalent cations at the time the IgG was added, and did not occur in the presence of the calcium ionophore A23187. Addition of aggregated IgG or chemotactic factors to lymphocytes thus may lead to a gated entry of calcium, and following closure of the calcium gate, the cells become relatively unresponsive to further stimulation.

Calcimycin↗
PubMed1978

Using pantographic tracings to detect TMJ and muscle dysfunctions.

Forty-six subjects were examined using the HDI and a PRI. Two sets of pantographic tracings were used to determine if subjects experiencing dysfunction could differentiate between degrees of dysfunction. Subjects were divided into groups according to their clinical symptoms: none (D0), sligh (D1), and moderate dysfunction (D5). Each subject was further classified into one of two groups according to his state of occlusion/articulation. The PRI detected differences between the group with moderate dysfunction and those groups with no dysfunction and slight dysfunction on the basis of differences between the first and second sets of tracings; no difference was found between the groups with slight and moderate dysfunction. Subjects with poor occlusions had higher PRI scores. The diagnosis of TMJ dysfunction may require the use of several modalities such as subjective responses, clinical examination, radiographs, and pantographic tracings.

Dental Articulators↗