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Biomedical subjects

C Wells

Publications and source records attributed to C Wells.

At least 127 records · Page 7Linked to original sources
PubMed1985

Fluorescence studies on the nucleotide- and Ca2+-binding domains of molluscan myosin.

The effects of nucleotides and Ca2+ on the intrinsic tryptophan fluorescence of molluscan myosin and its proteolytic fragments were studied. By using these proteins from the scallop, Pecten maximus, the existence of two distinct tryptophan-containing domains was established, which respond independently to ATP and Ca2+-specific binding. The latter is located in the 'neck' region of the myosin, which constitutes the regulatory domain. Subfragment 1, lacking the regulatory domain, responded only to ATP binding. On the other hand a tryptic fragment comprising the regulatory domain responded only to Ca2+ binding. Subfragment 1, containing the regulatory domain, responded to both ATP and Ca2+, but its ATPase activity was Ca2+-insensitive. By contrast, the ATPase activity of HMM was Ca2+-sensitive. Increasing the ionic strength had a detrimental effect on Ca2+-sensitivity, and fluorescence studies on solubilized myosin were therefore of limited value. Myosin and its fragments from other molluscan species which were investigated produced similar changes to those of Pectan maximus.

Adenosine Diphosphate↗
PubMed1985

Surgical and pathologic complications associated with peritoneovenous shunts in management of malignant ascites.

Forty-three peritoneovenous shunts have been inserted to palliate malignant ascites in 33 patients. Ascites was controlled for a time in every patient, but 18 shunts eventually blocked. Further shunt revision successfully controlled ascites until death in five of these patients and for prolonged periods in another five. The authors observed a marked difference between the performances of the two available shunts, but emphasize that the two groups of patients were not selected at random and therefore may not be comparable. Twelve postmortem examinations have been performed in the 33 patients to ascertain causes of shunt malfunction and to identify possible evidence of abnormal or accelerated tumor spread. The postmortem findings highlight great variability in the capacity of iatrogenically introduced showers of tumor cells to seed. There was a spectrum of tumor growth in the lung from a complete absence of tumor cells through dormant tumor clumps to developing metastases. The authors found no evidence either clinically or at autopsy, that the procedure had adversely affected the prognosis, except in one patient who died from pulmonary edema immediately after the operation.

Abdominal Neoplasms↗
PubMed1984

8-Anilino-1-naphthalenesulphonate, a fluorescent probe for the regulatory light chain binding site of scallop myosin.

Regulatory light chain (RLC) dissociation from scallop myofibrils, myosin or its subfragments was accompanied by an increase in binding of the hydrophobic fluorophore, 8-anilino-1-naphthalene-sulphonate (ANS) to the denuded proteins. The binding was monitored by the large increase in fluorescence emission at 460 nm when excited directly at 380 nm or via energy transfer from nearby tryptophan residues at 295 nm. ANS thus provides a convenient probe for following the kinetics of RLC dissociation in the presence of EDTA and its association in the presence of divalent metal ions. The observed RLC dissociation rate constant for myosin at 20 degrees C was 7.5 X 10(-3)S-1. The association rate constant, which was independent of the RLC concentration, was 5 X 10(-3) S-1. Subfragment 1, prepared by digestion of myosin in the presence of divalent metal ions to protect the light chains [S1(+LC)], showed reversible ANS binding qualitatively similar to the parent molecule. However when prepared in the presence of EDTA, subfragment 1 lacked RLC [S1(-LC)], its heavy chain molecular weight was reduced by about 4000 and it lacked the ANS binding region attributed to the RLC site. The tryptic digestion pattern of of S1(+LC) and S1(-LC) suggested that the 4000 difference peptide is at the C-terminus. Tryptic digestion of S1(+LC) has been shown to lead to the production of a regulatory peptide, comprising the two light chains and a heavy chain fragment, which displayed reversible ANS binding on addition of EDTA. Evidence is presented which suggests that this domain is at the C-terminus of subfragment 1.

Anilino Naphthalenesulfonates↗
PubMed1984

The characterization of vanadate-trapped nucleotide complexes with spin-labelled myosins.

The properties of spin-labelled myosin, prepared from rabbit skeletal and scallop adductor muscle, on forming a long-lived complex with ADP and vanadate (M.ADP.Vi), have been investigated. In the case of an iodoacetamide-based label attached to rabbit myosin or subfragment 1, M.ADP.Vi formation is characterized by a marked increase in the mobility of the probe, similar to that seen during steady-state ATPase activity. Hence, this complex appears to be a good analogue of the M**ADP.Pi state. The kinetics of M.ADP.Vi formation were determined by following the electron paramagnetic resonance (e.p.r.) signal with time and were analysed according to the scheme: (formula; see text) After correction for Vi polymerization, K'4 = 3.2 X 10(-4)M, k'-3 = 8.7 X 10(-3) s-1 and k'3 = 1.5 X 10(-4) s-1. The major effect of spin-labelling the reactive SH1 thiol is to increase k'3, so that M.ADP.Vi dissociates over a period of hours rather than days. In contrast, a maleimide-based spin-label attached to rabbit myosin does not exhibit a large change in mobility, on formation of the M.ADP.Vi complex. However, the small change observed in both the conventional and saturation transfer spectra questions the assumption that this probe is completely insensitive to librational motion during ATPase activity. The immobilized spectrum of the iodoacetamide-based spin label attached to scallop myosin is insensitive to M.ADP.Vi formation in the presence or absence of Ca2+. Under these conditions, the label appears to reflect gross head motion and hence this observation lends no support to the idea that, in the myosin-linked regulatory system, Ca2+ operates by controlling the flexibility of the subfragment 1-subfragment 2 joint.

Adenosine Diphosphate↗
PubMed1984

Electron microscopic evidence of antibody entry into neutrophils after phagocytosis of highly virulent group B streptococci.

An electron microscopic study was undertaken of the entry of specific antibody into neutrophils containing surviving intracellular highly virulent group B streptococci after phagocytosis of the organisms had occurred. Electron micrographs are presented to demonstrate that specific antibody gains access to the ingested bacteria. This antibody binds to the surface of the streptococci, which subsequently permits the neutrophils to kill these organisms.

Adult↗
PubMed1984

Interaction between Trichomonas vaginalis and other pathogenic micro-organisms of the human genital tract.

Trichomonas vaginalis organisms were mixed with suspensions of Neisseria gonorrhoeae, Mycoplasma hominis or Chlamydia trachomatis to allow ingestion of these micro-organisms by the trichomonads. Culture studies indicated that gonococci and mycoplasmas were ingested and that the number of intracellular viable organisms decreased rapidly, most gonococci being killed within six hours and all mycoplasmas within three hours. Electron microscopy revealed phagocytic uptake and destruction of these two micro-organisms within the trichomonads, gonococcal degradation being associated with lysosomal enzyme activity. There was no evidence from cultural or electron microscopy studies that C trachomatis organisms persisted in mixed culture with T vaginalis.

Chlamydia trachomatis↗
PubMed1983

Segmental flexibility and head-head interaction in scallop myosin. A study using saturation transfer electron paramagnetic resonance spectroscopy.

Saturation transfer electron paramagnetic resonance spectroscopy was used to investigate the rotational motion of the head domains of native and desensitized scallop myosin and its proteolytic subfragments. Scallop myosin was spin-labelled with 4-(2-iodoacetamido)-2,2,6,6-tetramethylpiperidinooxyl, which reacted with a heavy chain residue in the subfragment 1 domain. As previously shown for rabbit skeletal muscle myosin (Thomas et al., 1975), the two head domains of native scallop myosin appear to have independent motion (rotational correlation time, pi, = 0.8 X 10(-7) s for subfragment 1; 1.4 X 10(-7) s for myosin). However, removal of a regulatory light chain, to effect desensitization of the actin-activated ATPase, was associated with an increase in pi for myosin to a value of 2.4 X 10(-6) s. The Ca2+ sensitivity and initial correlation time were restored on recombination of the regulatory light chain in the presence of Mg2+. Sedimentation velocity profiles in an analytical ultracentrifuge indicated that the desensitized myosin preparations were largely monomeric and therefore the change in pi appears to reflect an intramolecular event. Addition of EDTA to spin-labelled scallop heavy meromyosin caused an immediate 2.5 to 4-fold increase in pi and a partial desensitization of the ATPase activity. Comparable experiments with subfragment 1 yielded a barely detectable increase in pi (1.5-fold) in the first ten minutes. The restricted rotational motion observed in desensitized myosin and heavy meromyosin could arise by a conformational change in the subfragment 1-subfragment 2 hinge region or by an association of one head with its partner. The latter mechanism, involving the exposed light chain binding site, would also explain the preferential release of one regulatory light chain from scallop myosin, and might account for some other co-operative effects observed in this molecule (Bagshaw, 1980).

Adenosine Triphosphatases↗
PubMed1983

Technical difficulties overcome in the use of Lowicryl 4KM EM embedding resin.

Several technical difficulties have been overcome in the use of Lowicryl 4KM resin. In order to embed and section tissue satisfactorily in the resin, it has been found necessary to thoroughly degass the resin before infiltration and polymerisation. After irradiation with UV light, the blocks are further polymerised by exposure to daylight for 2-3 weeks and then stored under partial vacuum over dessicant.

Acrylic Resins↗
PubMed1983

Epithelial patchy necrosis in Crohn's disease.

Electron and light microscopic studies of the intestinal epithelium in Crohn's disease demonstrated localized areas of damage to the superficial epithelium, occurring without accompanying acute inflammation. In a blind study of rectal biopsy specimens from seven patients with Crohn's disease, four with ulcerative colitis, and four normal controls, this finding of patchy necrosis without acute inflammation was observed exclusively in four of the seven cases of Crohn's disease.

Biopsy↗
PubMed1983

Randomised double-blind trial on the immediate effects of naloxone on classical Chinese acupuncture therapy for chronic pain.

The present study was designed to test the hypothesis that acupuncture therapy and acupuncture analgesia are both endorphin mediated. Thirty consecutive patients, all with pain at the time of treatment, were chosen. Classical acupuncture was used on seventeen patients; ear acupuncture was combined with classical Chinese acupuncture on eight patients, and ear acupuncture alone was used in five cases. No electrical stimulation of any sort was used. Pain was assessed by the patients on a visual scale of 0 to 100 before and after acupuncture therapy. Immediately following the post-acupuncture pain assessment, the patient was injected intravenously with a 1 ml. injection of 400 micrograms of Naloxone or a 1 ml. injection of 0.9% Sodium Chloride on a randomised double-blind basis. The effect on pain relief was noted immediately and after thirty minutes. No significant change in pain relief already obtained was noted after Naloxone in any patient. This does not support the hypothesis that acupuncture therapy is mediated by endorphin.

Acupuncture Therapy↗
PubMed1981

The use of concanavalin A crossed immuno-affinoelectrophoresis to detect hormone-associated variations in alpha 1-acid glycoprotein.

alpha 1-Acid glycoprotein produces three peaks on crossed immuno-affinoelectrophoresis with concanavalin A in the first dimension, which is indicative of a degree of heterogeneity in the carbohydrate portion of this plasma glycoprotein. A different pattern, however, showing relatively less concanavalin A-binding material, is found in association with and after an increase in female sex hormone levels. Our observations strongly suggest a prolonged hormonal effect on the carbohydrate composition of alpha 1-acid glycoprotein, which may alter the metabolism or the function of the protein.

Concanavalin A↗
PubMed1979

The mitogenic activity of lipopolysaccharide for spleen cells from germfree, conventional, and gnotobiotic rats.

Spleen cells from germfree rats, conventionally reared rats, and gnotobiotic rats associated with two Pseudomonas species gave no positive blastogenic response when incubated with each of four lipopolysaccharide (LPS) preparations from Escherichia coli, with glycolipid extracted from Salmonella minnesota R595 or with S. minnesota R595 lipid A. However, spleen cell preparations from athymic mice demonstrated a positive blastogenic response when incubated with E. coli LPS. Removal of adherent cells from germfree and conventional-flora rat spleen cells did not increase the mitogenic activity of LPS for nonadherent cells (less than 0.5% esterase-positive cells). All rat spleen cell preparations gave positive blastogenic responses to phytohemagglutinin and concanavalin A. This study indicates that LPS may not be a mitogenic agent for rat spleen cells.

Animals↗