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R Jones

Publications and source records attributed to R Jones.

At least 685 records · Page 38Linked to original sources

Identification of zona- and fucoidan-binding proteins in guinea-pig spermatozoa and mechanism of recognition.

Binding of guinea-pig spermatozoa to the zona pellucida of homologous eggs has been reported to involve 'receptors' on the inner acrosomal membrane (Huang et al. 1981). These receptors can be blocked by sulphated polysaccharides such as fucoidan (Huang and Yanagimachi, 1984). The aims of the present investigation were to identify these putative zona receptors using 125I-fucoidan as a probe and examine their mechanism of recognition. Results show that 125I-fucoidan binds to several proteins extracted from guinea-pig spermatozoa with molecular masses of 95, 60, 48, 34, 30 and 18-20 x 10(3) (K) on SDS-PAGE. The 48K, 34K and 30K components represent proacrosin and two forms of acrosin, respectively. 125I-zona pellucida glycoproteins also bound strongly to the 48K, 34K and 30K sperm proteins. The other high and low mass binding proteins were not positively identified but cytochemical experiments with fluoresceinamine-fucoidan and FITC-soybean trypsin inhibitor indicate that they are intraacrosomal. The mechanism of binding of 125I-fucoidan to proacrosin/acrosin (and also the 95K, 60K and 18K-20K components) involves multiple sulphate groups on the polysaccharide in a specific orientation to allow them to interact with basic residues on the protein. It is suggested that guinea-pig spermatozoa retain sufficient proacrosin/acrosin bound to the inner acrosomal membrane after the acrosome reaction to mediate binding to the zona pellucida and that functionally proacrosin is analogous to sea urchin binding.

Acrosin↗

Spatial organization of repetitive DNA sequences in the bovine sperm nucleus.

During spermatogenesis, DNA in the sperm head becomes more tightly condensed as histones are replaced by protamine-like molecules. In this article, the question is asked whether, during the production of this highly differentiated cell, controls are imposed on the spatial organization of DNA within the nucleus. Heads from bull spermatozoa were isolated by a technique that removed the plasma membrane and acrosomal contents, and the DNA was induced to decondense by addition of 2-mercaptoethanol and trypsin. Under these conditions, decondensation was induced in all regions of the head. To determine whether there was any spatial restraint on packaging of the genome, three DNA probes were used (pl.709-512, containing an interspersed repetitive sequence; pCSIH, containing a copy of the major bovine centromeric statellite sequence; p18 s and p28 s, containing the 18 S and 28 S ribosomal genes) that might be expected to hybridize to different regions. Results showed that the interspersed repetitive probe hybridized to all regions of the head, whereas the ribosomal and centromeric probes hybridized to sequences that were largely confined to the equatorial region of the sperm. We conclude that organization of the genome in the bovine sperm nucleus is not random.

Animals↗

Density-gradient separation for 4-hydroperoxycyclophosphamide purging of autologous bone marrow grafts.

We studied density-gradient separation of autologous bone marrow grafts in preparation for ex vivo 4-hydroperoxy-cyclophosphamide (4-HC) purging. A two-step procedure of buffy coat isolation followed by Ficoll-diatrizoate separation was used. Buffy-coat cells were isolated primarily either using the COBE 2991 Blood Cell Processor or Haemonetics 30 Cell Separator. All density-gradient separations were performed using the COBE 2991. The nucleated cell recoveries after buffy-coat isolation were 44.3 +/- 10.4% and 85.1 +/- 10.5% (+/- standard deviation, P less than 0.001) for the Haemonetics 30 and COBE 2991 isolated grafts, respectively. The final nucleated cell recoveries after density-gradient separation for these two buffy-coat techniques were 24.2 +/- 10.1% and 29.5 +/- 14.7% (p = 0.30). Compared to manual density-gradient separation, the techniques using the COBE 2991 recovered slightly higher numbers of nucleated cells and CFU-GM. The red blood cell concentration of the 4-HC incubation mixture was 1.0 +/- 0.6% and the CFU-GM survival after treatment was 2.4 +/- 2.6%. The variability in CFU-GM survival after 4-HC incubation of the density-gradient separated grafts appears less than previous experience treating buffy-coat cells. Density-gradient separation of autologous bone marrow appears to provide a more uniform treatment with 4-HC and may improve the clinical transplant course of the autograft recipient.

Bone Marrow↗

Horizontal fusional amplitudes after adaptation to prism.

Horizontal fusional limits and lateral phorias were measured subjectively on five normal subjects after adaptation to increments of base-in and base-out prism. Both the phoria and the fusional limits were found to change by approximately the amount of the adapting prism. The fusional amplitudes (the difference between the phoria and the fusional limits) were relatively unaffected by vergence adaptation except at the largest values of adapting prism. These results are in agreement with current models of disparity vergence eye movement control which suggest that there are independent fast and slow fusional vergence mechanisms, each with its own saturation limit.

Adaptation, Ocular↗

Do women and myopes have larger pupils?

It is commonly reported that the pupils of women are larger than those of men and the pupils of myopes larger than those of emmetropes. However, these reports are not supported by experimental procedures using objective measurements, controlled conditions, and adequate numbers of subjects. In this report the open-loop pupil size was measured objectively using dynamic infrared pupillometry in a sample of 48 subjects. Subjects were balanced for age and equally divided between emmetropes and myopes, males and females. The results did not support the contention that females have larger pupils than males or that physiologic myopes have larger pupils than emmetropes.

Adolescent↗

Ventricular performance and glucose uptake in rats during chronic hypobaric hypoxia.

To identify changes of ventricular performance and their relationship to myocardial glucose uptake in Sprague-Dawley rats exposed to hypobaric hypoxia, radionuclide angiocardiograms (n = 34) and 2-[14C]deoxyglucose (2-[14C]DG) autoradiography (n = 14) were performed on rats maintained either for two weeks in air at 380 mmHg (hypoxic group), two weeks in hypobaric hypoxia followed by two weeks of air (recovered group), or in air (control group). Right ventricular ejection fraction (RVEF) was 66% +/- 2% (mean +/- s.e.m.) in controls, 40% +/- 3% during hypoxia, and 60% +/- 2% in recovered rats. LVEF remained unchanged. In controls, RV 2-[14C]DG uptake was 77% +/- 3% of LV uptake. During hypoxia, 2-[14C]DG uptake increased. This increase was greater within the RV than the LV and septum (85 +/- 16% versus 51 +/- 10%, p less than 0.005). The alterations of RV 2-[14C]DG uptake correlated with systolic pulmonary artery pressure (r = 0.77, p = 0.002).

Animals↗

Prevention of neutrophil-mediated injury to endothelial cells by perfluorochemical.

Myocardial salvage after reperfusion may be limited by neutrophil-mediated microvascular damage. The effect of the perfluorochemical, Fluosol-DA, and its various components on neutrophil adherence, cytotoxicity, and proteolytic enzyme release was examined on sheep large and small vessel endothelial cells in vitro. Cells were studied under normoxic (N) and anoxic conditions (A). Various concentrations of Fluosol (10%, 25%, and 50%) significantly reduced neutrophil adherence under both experimental conditions [mean 22 +/- 3.25% versus 7 +/- 0.8% (N) and 20 +/- 3.2% versus 7.5 +/- 0.9% (A); P less than 0.01]. The perfluorocarbons, perfluorodecalin (PFD), and perfluoro-tripropylamine (PFTP) in a 50 volume/percent concentration exhibited profound effects on adherence, particularly on cells subjected to anoxia (51% and 69% reduction in adherence, respectively; P less than 0.01). No effect on adherence was observed with other components, including the detergent, pluronic F68. A 25% reduction (P less than 0.02) in endothelial cytotoxicity was noted when neutrophils were preincubated with Fluosol. However, pretreatment of endothelial cells with Fluosol did not inhibit neutrophil adherence. Neutrophils stimulated with cytochalasin B and FMLP showed a significant reduction in lysozyme release after incubation with Fluosol (28 +/- 5% versus 17 +/- 4%; P less than 0.01). This study demonstrates that Fluosol significantly attenuates neutrophil adherence, cytotoxicity, and enzyme release in an in vitro model of microvascular injury. It also suggests that prevention of neutrophil-mediated microvascular damage may be an important mechanism whereby Fluosol enhances myocardial salvage after ischemia and reperfusion.

Animals↗

Irritable bowel syndrome--making the diagnosis.

Irritable bowel syndrome need not be a diagnosis of exclusion. It can be made with confidence on clinical grounds, without wasting secondary care resources or increasing the patient's anxiety by unnecessary investigations.

Abdominal Pain↗

High-dose, double alkylating agent chemotherapy with DTIC, melphalan, or ifosfamide and marrow rescue for metastatic malignant melanoma.

Thirty-seven patients with widely metastatic malignant melanoma were treated with one of three chemotherapy regimens, incorporating high-dose dacarbazine (DTIC). The chemotherapy was followed by autologous bone marrow rescue which was harvested under local anesthesia in 25 of the patients. The three regimens comprised a 24-hour infusion of DTIC (Regimen A for patients less than 45 years of age, 4.3 to 10.5 g/m2; B, if greater than 45 years of age 2.7 to 4.0 g/m2; and later C, if greater than 45 years of age 7.0 to 8.0 g/m2). The second alkylating agent was given at +8 and +16 hours from the start of DTIC. The total doses of the melphalan ranged from 60 to 130 mg/m2 for Regimen A and 30 to 40 mg/m2 for Regimen B. Ifosfamide 5.0 to 8.0 g/m2 was given instead of melphalan in Regimen C. The response rates for the regimens were 81% (25% CR) for A, 27% (11% CR) for B, and 20% (with no complete responders) for Regimen C. There was no statistically significant difference between the three regimens for survival with a median value of 6 months. One of the 16 patients treated with the very high dose Regimen A died of septicemia and three of ten patients in Regimen C died within the first 2 weeks of treatment. There was statistically significant greater myelosuppression, stomatitis, and diarrhea in the very high dosage DTIC and melphalan (Regimen A) compared with the other two regimens. No significant difference in response rate or toxicity was observed for the different dosages escalated within each of the three regimens. Although hematologic and gastrointestinal toxicity were very severe, no unusual side effects were noted except for one episode of severe acute renal failure in the high-dose DTIC and melphalan, Regimen A. Responses occurred mainly in nonvisceral, nodal, and cutaneous sites and occasionally in pulmonary metastases. The Karnofsky performance improved 4 to 6 months after treatment notably with the high-dose DTIC and melphalan therapy. No survival benefit for the combination chemotherapy despite the high dosages was detected and such an approach currently cannot be recommended.

Adult↗

Poly(ADP-ribosylation) of nuclear proteins in rat testis correlates with active spermatogenesis.

Poly(ADP-ribosylation) of nuclear proteins has been investigated in rat testis under different experimental conditions to determine whether it is associated with somatic or germinal cells. Isolated, intact nuclei were incubated with [14C]NAD and extracted sequentially with 5% HClO4 and 0.25 M HCl, and labelled soluble proteins were analysed by reverse-phase high-performance liquid chromatography and acetic acid-urea polyacrylamide gel electrophoresis (pH 2.9). Results show that in normal adult testis a major acceptor protein for poly(ADP-ribose) in HClO4 extracts is the tissue-specific histone, H1t. Core histones and three proteins (alpha, beta and gamma) with low mobility on acetic acid-urea gels were the major acceptors identified in HCl extracts. Poly(ADP-ribosylation) of all the aforementioned proteins is very low in isolated intact nuclei of testis from 8-day-old animals (only spermatogonia present in seminiferous tubules), increases significantly by 16-day (pachytene spermatocytes appear) and reaches adult proportions by 32 days (condensing spermatids present). In the nuclei from cryptorchid testes, poly(ADP-ribosylation) of nuclear proteins resembles 8-day-old testis. It is concluded that (a) poly(ADP-ribosylation) of nuclear proteins in rat testis is closely correlated with spermatogenesis and can be inferred that is particularly active in the early stages of meiosis; (b) testis-specific proteins (histone H1t and low mobility proteins, alpha, beta and gamma) are poly(ADP-ribosylated) to higher specific radioactivity than somatic histones.

Age Factors↗

Improvement of ischemia-reperfusion-induced myocardial dysfunction by modulating calcium-overload using a novel, specific calmodulin antagonist, CGS 9343B.

The present paper explores the mechanism of calcium-overloaded cardiac cell exocytosis during reperfusion of ischemic myocardium. A novel specific inhibitor of calmodulin, CGS 9343B, was used to pretreat an ischemic heart in an effort to enhance myocardial preservation. The experimental model employed an isolated in situ pig heart subjected to 120 min of ischemic insult by reversibly occluding the left anterior descending coronary artery, the last 60 min being superimposed with global hypothermic cardioplegic arrest. This ischemic episode was followed by 60 min of revascularization. CGS 9343B enhanced post-ischemic myocardial recovery, as judged by improved regional as well as global myocardial functions, better preservation of high-energy phosphate compounds, and reduced release of creatine kinase. Since this compound blocks calmodulin without inhibiting protein kinase C, the results of this study suggest that calmodulin-dependent kinase, rather than protein kinase C, is primarily involved in expressing calcium-overloaded cell exocytosis, and a specific calmodulin antagonist such as CGS 9343B can be used to salvage an ischemic heart from reperfusion injury.

Animals↗

Prevalence of symptoms of dyspepsia in the community.

To study the prevalence of dyspepsia in the community a postal questionnaire was sent to 2697 patients who were selected at random from the lists of patients registered in two health centres in Hampshire. A total of 2066 returned questionnaires were suitable for analysis (response rate 77%). It was found that the six month prevalence of dyspepsia was 38%. There was considerable overlap between symptoms of heartburn and upper abdominal pain, with over half of patients with dyspepsia experiencing both. One in four of these patients had consulted their general practitioner during that time. The proportion of patients with dyspepsia who consulted their general practitioner varied widely among the eight doctors who participated in the study, from 17% to 45%. Frequency of symptoms tended to fall with age, particularly in men, while the proportion of patients with dyspepsia who sought medical advice increased with age. Almost one in five of the 2066 patients had been investigated with radiology or endoscopy at some time, and 143 (7%) of them claimed to have had a diagnosis of peptic ulcer.

Adult↗

Gaze distractibility in Wilson's disease.

We describe the case of a patient with Wilson's disease who was first seen with distractibility of gaze fixation. Clinical and magnetic resonance imaging studies suggest that the disorder was probably due to frontal lobe involvement.

Adult↗