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Biomedical subjects

J Underwood

Publications and source records attributed to J Underwood.

At least 19 recordsLinked to original sources
PubMed1991

The relationship between the light scattering properties of zymogen granules and the release of their contained proteins.

The optical density of suspensions of the digestive enzyme-containing zymogen granule, a roughly spherical 1 micron diameter membrane-enclosed subcellular structure isolated from the exocrine pancreas of mammals, is reduced greatly when they are suspended in physiological media. This reduction in optical density is accompanied by the release of the granule's protein contents. It has traditionally been assumed that this property is due to granule lysis; that is, dissolution of the particle and its consequent disappearance as a strongly scattering object. Thus, lysis would decrease optical density by decreasing the number density of suspended spheres (N) according to Beer's law. However, as a general matter, changes in the optical density of suspensions of spheres may be a function of changes in the refractive index (m) or radius (r) of the objects as well. In this study, we apply Mie theory of scattering by small particles, which, in conjunction with Beer's law, allows us to evaluate whether changes in the scattering properties of granule suspensions are due to changes in N, m or r. Scattering by granule suspensions was reduced in three ways-pH, calcium ion concentration, and detergent concentration. A simple reduction in particle number did not account for decreased scattering and protein release in any of these circumstances. Instead, the changes appear attributable to decreases in particle size and refractive index.

Animals↗
PubMed1989

High-performance liquid chromatography of amino acids, peptides and proteins. XCIII. Comparison of methods for the purification of mouse monoclonal immunoglobulin M autoantibodies.

A comparison of methods for the purification of naturally occurring mouse monoclonal autoantibodies, of the immunoglobulin M (IgM) isotype, has been performed to determine the optimal strategies for the isolation of IgM from ascites fluid and in vitro tissue culture hybridoma supernatants. In order to quantify each purification procedure, the concentration of IgM in eluted fractions was determined by using a double-sandwich mu-chain-specific anti-IgM enzyme-linked immunosorbent assay, and the purity of the IgM was determined by a bicinchoninic acid-based protein assay and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The most efficient single-step purification was based on size-exclusion chromatography on high-resolution Superose 6 HR 10/30 fast protein liquid chromatography (FPLC) columns. This procedure resulted in recoveries of monoclonal IgMs of ca. 71-86% with purities between 68 and 86%. Single-step chromatography of monoclonal IgM, on Superose 6 FPLC columns resulted in a 21-fold purification of IgM, prepared by the in vitro culture of hybridoma cells in dialysis membrane. Size-exclusion chromatography, performed with Sephacryl S-300 columns, resulted in reduced resolution of monoclonal IgM, with yields of ca. 57-80% and purity of ca. 42-58% compared with the high-resolution Superose 6 FPLC columns. "Non-ideal" size-exclusion chromatography on Superose 6 FPLC columns resulted in selective retention of monoclonal IgMs and elution of IgM with high-ionic-strength buffers in the trailing peak. Recovery of IgM with this strategy was high (ca. 82-92%) but the purity was not comparable to the single-step fractionation of IgM on Superose 6 FPLC columns. Single-step anion- and cation-exchange and mixed-mode hydroxyapatite chromatography resulted in only partial purification of monoclonal IgM with the applied procedures. With these latter separation techniques, monoclonal IgM was eluted with a variety of other ascites fluid or supernatant proteins, including those with apparent molecular weights identical to those of mouse IgG and albumin. Sequential purification of monoclonal IgMs by Mono Q anion exchange, followed by Superose 6 FPLC columns, resulted in a 2- to 3-fold purification of IgM but did not separate IgM from high-molecular-weight contaminants with apparent molecular weights similar to those of alpha 2-macroglobulin and IgG. Enrichment of monoclonal IgM from ascites fluid by ammonium sulphate precipitation revealed increasing IgM recovery with increasing ammonium sulphate final concentrations up to 60%.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acids↗
PubMed1989

Immunosuppressive activity in human in vitro fertilization (IVF) culture supernatants and prediction of the outcome of embryo transfer: a multicenter trial.

The supernatants from cultured human oocytes fertilized in vitro contain low molecular weight factors that can suppress or stimulate the proliferative response of lymphocytes in vitro. The inhibitory and stimulatory effects are nonspecific and may be detected using cultured human or murine tumor cell lines. Using such a bioassay, we previously tested fetal cord serum-supplemented culture supernatant and found that an absence of suppression was correlated with an absence of subsequent pregnancy. To test this association further, additional samples were obtained from four different in vitro fertilization (IVF) units and studied blindly without knowledge of the pregnancy outcome. In this series, samples were obtained after the first 12-24 hr of sperm-oocyte incubation and all of the supernatants were from individual embryo cultures. The average number of preembryos transferred to those achieving pregnancy did not differ significantly from the number transferred to those not achieving pregnancy but the level of suppression was greater (8.7 +/- 1.9%) in those becoming pregnant compared to those not achieving pregnancy (0.8 +/- 1.5%). Twenty-two of 61 patients who received at least one embryo with a suppressive supernatant achieved pregnancy, whereas 0 of 19 patients received embryos lacking suppressive supernatants became pregnant. Two patients who received a single embryo from cultures with suppression became pregnant. Several problems with the bioassay method were defined. The culture medium in this series was always supplemented with adult serum, usually from the patient herself, and this serum could be suppressive.(ABSTRACT TRUNCATED AT 250 WORDS)

Blastocyst↗
PubMed1989

Histologic and immunologic study of uterine biopsy tissue of women with incipient abortion.

Biopsy specimens taken from the region of the placental bed were examined for the presence of phloxinophilic granulated mononuclear cells in women with a history of recurrent miscarriage and who would eventually miscarry a current pregnancy. They were compared with biopsy specimens from women with intact pregnancies presenting for elective termination of pregnancy and those with "missed abortion." Cells with large cytoplasmic granules (greater than or equal to 1 micron) were abundant in the group of ongoing pregnancies whereas cells with smaller granules (less than 1 micron) that were similar to large granular lymphocytes were more abundant relative to cells with large granules in the biopsy specimens from failing pregnancies. Immunosuppressive activity was tested in the supernatants of cultured biopsy samples of each group and found to be significantly lower in the incipient miscarriage group. These findings could represent alterations associated with the process of miscarriage, such as inflammation, or there may be deficient suppressor cell activity at the fetomaternal interface as the reason for "rejection" of the early embryo.

Abortion, Habitual↗
PubMed1987

Histopathologic alterations in the decidua in human spontaneous abortion: loss of cells with large cytoplasmic granules.

The histopathology of decidua obtained from the placental bed was evaluated by phloxine-tartrazine staining, which allows clear definition of cells with cytoplasmic granules. Mononuclear cells with large granules were seen in biopsy specimens taken from women at 8-31 weeks of normal pregnancy. In contrast, cells with large granules were missing in sections taken from the decidua of five women who were aborting or were destined to abort. Since the presence of suppressor cell activity in murine decidua correlates with the success of pregnancy and since this suppression is associated with small lymphocytes with cytoplasmic granules, the observations made using human placental bed biopsy material suggest that a possible suppressor cell deficiency might occur in the early stages of spontaneous abortion in human females.

Abortion, Spontaneous↗
PubMed1977

Simple hypothermic preservation for transporting human livers long distances for transplantation. Report of 12 cases.

Since 1973, 12 patients in the Cambridge-King's College Hospital liver transplant programme have received livers from donors dying in hospitals considerable distances from the transplant centre in Cambridge. The method of preservation used to transport these livers from 35 to 110 miles was flush perfusion with plasma protein fraction and hypothermic storage in ice. The ischaemia times ranged from 2 hr and 42 min to 4 hr and 22 min. All of the recipients had good or excellent postoperative function of the grafts and 6 of the 12 recipients are alive, the longest survival being 29 months. The preservation technique is simple and easily portable and has given reliable 4-hr preservation of the human liver.

Adolescent↗