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Biomedical subjects

M Cochran

Publications and source records attributed to M Cochran.

At least 37 records · Page 2Linked to original sources

Longevity of glass-ionomer restorative materials: results of a 10-year evaluation.

Three restorative materials used for cervical erosion /abrasion lesions were evaluated clinically after 10 years. Thirty adult patients with at least four cervical lesions received one restoration of each of Ketac-Fil, finished immediately, Ketac-Fil, finished after a delay, Chelon-FIl (all glass-ionomer cements), and Cervident (a resin composite). Restorations were placed without any tooth preparations. Eighteen patients returned for their 10-year examinations. Two experienced examiners provided the periodic evaluations. Complete retention at 10 years was 83% for Ketac-Fil, finished immediately; 78% for Ketac-Fil, finished after a delay; 67% for Chelon-Fil; and 17% for Cervident. All three glass-ionomer restorative materials exhibited statistically significantly greater retention that did Cervident. When a noninvasive procedure is desired, glass-ionomer materials are the restorative material of choice for abrasion/erosion lesions because of their long-term retention values.

Adult↗

Comparison of bone formation rates measured by radiocalcium kinetics and double-tetracycline labeling in maintenance dialysis patients.

We report 23 prospective studies on 18 maintenance dialysis patients in whom we measured skeletal mineralization rate (m) using 47Ca, analyzed by the expanding pool model, and compared it with the histologic bone formation rate (bfr), volume referent, estimated on tetracycline-labeled iliac crest bone. The patients showed a spectrum of bone disease types including adynamic bone, aluminum-related osteomalacia, and various degrees of secondary hyperparathyroidism. The mean width between double labels, on which mineral apposition rate depended, was estimated using a simple formula relating area to perimeter for each feature enclosed by the labels. Values for m ranged from 0 to 155 mmol calcium per day and for bfr from 0 to 124% per year. There was close correlation between m and bfr (r = 0.976), serum alkaline phosphatase (r = 0.968), and serum immunological parathyroid hormone (iPTH) (r = 0.868). When the volumetric bfr was converted to mass units and applied to the whole skeleton, using literature values for mineral density and cortical and trabecular mass, there was close agreement between the histologic and isotopic estimates of m (r = 0.959). The results validate the two methods and suggest they are interchangeable. However, use of a rigorous method to determine bfr appears to be essential.

Adult↗

Effect of citrate infusion on urinary aluminium excretion in the rat.

1. Seven pairs of rats received 1 mmol/l aluminium citrate in their drinking water 5 days before the experiments. Five additional rats were treated identically. Six rats received the same food but drank distilled water. 2. After a 6 h fast, the animal was anaesthetized, the jugular vein and femoral artery were cannulated and the bladder was catheterized, after which an intravenous infusion of Hartmann's solution containing [14C]inulin was begun. The urine was collected at 20 min intervals and 1 ml of arterial blood was obtained before the end of each collection. After at least two basal collections, the infusion was modified to contain, in addition, 5 mmol/l NaHCO3 (control) or 5 mmol/l sodium citrate (experimental). The infusion rate, constant in each pair, differed between pairs across the range 60-125 mumol/min. 3. A total of eight collections was made per animal and urine flow, glomerular filtration rate, plasma and urinary aluminium and citrate were measured. 4. Control and experimental rats had a higher mean basal plasma aluminium level (0.39 +/- 0.21 mumol/l) than the six rats receiving distilled water (0.16 +/- 0.14 mmol/l, P < 0.001). The corresponding urinary aluminium excretion rates were similar (46 +/- 31 and 47 +/- 23 pmol/min, respectively). There was no significant difference between the basal values of any variable in the control and experimental rats. No significant change was observed in any variable during the infusion of NaHCO3 (controls). Among the experimental rats, there was no significant change in urine flow, glomerular filtration rate or plasma aluminium level.(ABSTRACT TRUNCATED AT 250 WORDS)

Aluminum↗

A comparison between estimates of GFR using [99mTc]DTPA clearance and the approximation of Cockcroft and Gault.

BACKGROUND: The formula of Cockcroft and Gault, relating age, weight, sex and plasma creatinine is widely used to estimate glomerular function. Its ability to predict [99mTc]DTPA clearance, an accepted measure of GFR, is therefore of interest. AIM: To test prospectively the reliability of the formula in predicting the [99mTc]DTPA clearance in subjects with normal and sub-normal renal function. METHODS: The 24 hour creatinine clearance (Ccr), the derived clearance using the formula (Cder), and the [99mTc]TPA clearance have been compared in 47 male and 47 female subjects, aged between 15 and 71 years. Obese and non-obese subjects (males > 90 kg, females > 80 kg body weight) were considered separately. RESULTS: Across the range of [99mTc]DTPA clearance studied (14 to 125 mL/minute), in non-obese subjects, both Ccr and Cder were correlated similarly with [99mTc]DTPA clearance in males and females. In obese subjects as a group, Cder was also closely correlated with [99mTc]DTPA clearance but the scatter of data was wider. Among non-obese subjects, correlations were high between either Ccr or Cder and [99mTc]DTPA clearance values < 101 mL/minute but were not significant at [99mTc]DTPA clearance values > 100 mL/minute. For [99mTc]DTPA clearance values between 14 and 100 mL/minute, Cder was a more direct and more precise predictor of [99mTc]DTPA clearance [DTPA clearance = 1.01 (Cder) + 2.34, with 95% confidence limits of +/- 18 mL/minute] than of Ccr [Ccr = 1.34(CderC)-10.86, confidence limits +/- 28 mL/minute]. CONCLUSION: For very mild through to moderate renal failure, the formula of Cockcroft and Gault can evidently be used as a reasonable estimate of GFR.

Adolescent↗

Absorbed aluminium is found with two cytosolic protein fractions, other than ferritin, in the rat duodenum.

After in vivo perfusion of the upper intestine of the rat with a range of concentrations of aluminium chloride, entry of the metal into the portal system was only detected when the perfusate exceeded 400 mumol/l, suggesting a mucosal block. Using gel filtration of a mucosal cytosol extract, two consistently appearing aluminium peaks were identified which may represent aluminium binding proteins. Both were heat stable at 60 degrees C and had molecular sizes of about 700 (kilo daltons) (kD) and 17 kD respectively. The larger molecule was distinct from ferritin. Neither molecule associated with 59Fe nor 45Ca. It is suggested that the aluminium peaks are relatively specific aluminium binding proteins that have a scavenging role, reducing entry of the metal from the intestinal contents into the portal blood.

Aluminum↗

Interleukin-11 promotes accessory cell-dependent B-cell differentiation in humans.

Interleukin-11 (IL-11) is a recently described stromal-derived cytokine that supports the growth of an IL-6-dependent murine plasmacytoma line in the presence of antibody to IL-6 and appears to act in a manner similar to IL-6 on hematopoietic stem cells. Because IL-6 is known to promote differentiation of normal human B cells, the role of IL-11 on B-cell differentiation in vitro was characterized. IL-11 does not result in significantly increased DNA synthesis or Ig secretion by purified B cells alone or B cells cultured with Staphylococcus Cowan I, a T-cell-independent B-cell mitogen. In contrast, purified B cells cultured in the presence of pokeweed mitogen (PWM), irradiated T cells, and monocytes show increased DNA synthesis at day 3 and increased IgG and IgM secretion at day 7 of culture; addition of IL-11 further augments Ig secretion without change in DNA synthesis, an effect that can only be partially blocked by monoclonal antibody to IL-6. Similar experiments confirmed that increased IgG secretion was demonstrable when either IL-11 or IL-6 was added to B cells + CD4+/45RA- T cells + monocytes + PWM; in contrast, Ig secretion was low and equivalent when CD4+/45RA+ T cells were cultured with B cells+monocytes+PWM with or without IL-6 or IL-11. Neither IL-6 nor IL-11 could significantly increase phytohemagglutinin (PHA)-induced DNA synthesis by CD4+/45RA- or CD4+/45RA+ T cells. Although PWM or IL-11 induced IL-6 mRNA expression in both CD4+/45RA- T cells and monocytes, in neither cell did IL-11 increase IL-6 mRNA expression over that noted to PWM alone. These observations support the view that IL-11 promotes differentiation of human B lymphocytes only in the presence of accessory T cells and monocytes and that a minor component of this effect may be through stimulation of IL-6 production by CD4+/45RA- T cells and monocytes.

Actins↗

Safety and immunogenicity in volunteers of a recombinant Plasmodium falciparum circumsporozoite protein malaria vaccine produced in Lepidopteran cells.

A recombinant Plasmodium falciparum circumsporozoite (CS) antigen (rPfCSA) was produced in insect cells using a baculovirus expression vector containing the entire CS gene. This near full-length CS antigen was adsorbed onto aluminium phosphate for use as a malaria vaccine. In a study of safety and immunogenicity, 20 volunteers were divided into four groups of five each and inoculated intramuscularly with 10, 100, 500 or 1000 micrograms of vaccine. Primary vaccinations were followed by two booster immunizations at 2 and 6 months. Three volunteers developed prominent local reactions manifested as tenderness, redness and swelling at the injection site following the second or third vaccination. All symptoms resolved spontaneously within 72 h. Postimmunization sera from six of 20 volunteers showed seroconversions as measured by Western blot, using rPfCSA as antigen. However, specific anti-CS protein antibody could not be detected by indirect immunoflourescence against intact sporozoites or by ELISA using rPfCSA or peptide to the repeat region. In addition, 18 of 20 volunteers developed antibody to baculovirus proteins as determined by ELISA and/or Western blot. Antigen-driven replication studies using peripheral blood mononuclear cells from vaccinees failed to detect proliferative responses specific to CS protein. This recombinant CS protein vaccine, as formulated, was minimally immunogenic in humans.

Adult↗

Iron uptake by human reticulocytes at physiologic and sub-physiologic concentrations of iron transferrin: the effect of interaction with aluminum transferrin.

We have studied the interaction, in vitro, between diferric transferrin (FeTr), aluminum transferrin (AlTr), and human reticulocytes harvested from human placental blood. In particular, we aimed to determine the extent to which the kinetics of AlTr and FeTr differed. Using transferrin labeled with either 59Fe or 125I, the association of radiotracer with reticulocytes, as a function both of time and of transferrin concentration, was examined. Under the conditions of the experiments, the data are consistent with a mechanism involving at least three processes. Two early processes acting in parallel behave as a high-affinity saturable receptor and a low-affinity non-saturable receptor, neither of which distinguish between AlTr and FeTr. In a subsequent process, AlTr and FeTr exhibit different kinetics. This third process may be saturated by FeTr but not by AlTr. Interpreted in terms of a current conventional view of metallo-transferrin uptake, we conjecture that the early parallel processes involve cell surface phenomena including classical transferrin-receptor binding, and that the subsequent process represents events, possibly intracellular, involved in metallo-transferrin dissociation or further iron transport. The extent to which AlTr influences the interaction of FeTr with reticulocytes offers insight into both the normal physiology of iron uptake and the potential for toxicity by aluminum.

Binding, Competitive↗

HCO3(-)-coupled Na+ influx is a major determinant of Na+ turnover and Na+/K+ pump activity in rat hepatocytes.

Recent studies in hepatocytes indicate that Na(+)-coupled HCO3- transport contributes importantly to regulation of intracellular pH and membrane HCO3- transport. However, the direction of net coupled Na+ and HCO3- movement and the effect of HCO3- on Na+ turnover and Na+/K+ pump activity are not known. In these studies, the effect of HCO3- on Na+ influx and turnover were measured in primary rat hepatocyte cultures with 22Na+, and [Na+]i was measured in single hepatocytes using the Na(+)-sensitive fluorochrome SBFI. Na+/K+ pump activity was measured in intact perfused rat liver and hepatocyte monolayers as Na(+)-dependent or ouabain-suppressible 86Rb uptake, and was measured in single hepatocytes as the effect of transient pump inhibition by removal of extracellular K+ on membrane potential difference (PD) and [Na+]i. In hepatocyte monolayers, HCO3- increased 22Na+ entry and turnover rates by 50-65%, without measurably altering 22Na+ pool size or cell volume, and HCO3- also increased Na+/K+ pump activity by 70%. In single cells, exposure to HCO3- produced an abrupt and sustained rise in [Na+]i from approximately 8 to 12 mM. Na+/K+ pump activity assessed in single cells by PD excursions during transient K+ removal increased congruent to 2.5-fold in the presence of HCO3-, and the rise in [Na+]i produced by inhibition of the Na+/K+ pump was similarly increased congruent to 2.5-fold in the presence of HCO3-. In intact perfused rat liver, HCO3- increased both Na+/K+ pump activity and O2 consumption. These findings indicate that, in hepatocytes, net coupled Na+ and HCO3- movement is inward and represents a major determinant of Na+ influx and Na+/K+ pump activity. About half of hepatic Na+/K+ pump activity appears dedicated to recycling Na+ entering in conjunction with HCO3- to maintain [Na+]i within the physiologic range.

Animals↗

Cross sectional and longitudinal study of bone mineral content of the distal forearm in adult premenopausal women.

The range of fat corrected distal forearm mineral content (FMC) (at a site which comprises approximately 75% cortical and 25% trabecular bone) and the effect of aging on FMC were determined in normal premenopausal women. In 106 women (mean age 39; range 18-56) the mean FMC was 1.184 (SD 148) g/cm; fat correction increased this to 1.239 (SD 148) g/cm. There were significant correlations between fat corrected FMC and height, weight and BMI (r greater than 0.16; P less than 0.05). In 43 of these women (mean age 35; range 18-51) repeat measurements of FMC were performed. The last measurement was separated from the first by a mean time interval of 27 +/- 3 months. There was no significant change in uncorrected FMC (baseline 1.185 +/- 0.21 g/cm compared with their final measurement 1.182 +/- .020 g/cm; P = 0.51). Fat-corrected FMC (baseline 1.229 +/- .021 g/cm compared with 1.223 +/- .020 g/cm later) tended to decrease although this change was not significant (P = 0.06). Age ranked cusum analysis demonstrated a non-significant fat-corrected rate of change in FMC of -.3% yr in those over 35 years of age (P less than 0.10). This data suggests that if there is any distal forearm bone loss before the menopause it is negligible.

Adolescent↗

Plasma membrane H(+)-HCO3- transport in rat hepatocytes: a principal role for Na(+)-coupled HCO3- transport.

Na(+)-coupled HCO3- transport has been demonstrated in the basolateral membrane of hepatocytes, but there is uncertainty regarding its stoichiometry or capacity compared with other mechanisms of H(+)-HCO3- transport. After preincubation in medium free of Na+, either in the presence or absence of HCO3(-)-CO2, rat hepatocytes in primary culture were reexposed to Na+ or HCO3(-)-CO2 alone or in combination. Transporter electrogenicity was assessed by measuring membrane potential difference (PD), and the relative capacities of Na(+)-coupled HCO3- transport, Cl(-)-HCO3- exchange, and Na(+)-H+ exchange were assessed by measuring the magnitude and rate of change of intracellular pH (pHi) using BCECF. In the absence of Na+, exposure to HCO3- alone had no consistent effect on PD or pHi. In the absence of HCO3-, reexposure to Na+ depolarized cells by 3 +/- 1 mV and caused an amiloride-inhibitable increase in pHi of 0.031 +/- 0.02 units/min. In the presence of HCO3-, reexposure to Na+ hyperpolarized cells by -14 +/- 5 mV and increased pHi at a rate of 0.133 +/- 0.11 units/min; both the hyperpolarization and alkalinization were inhibited by SITS but unaffected by amiloride. These changes in PD indicate that Na(+)-coupled HCO3- transport is electrogenic, consistent with coupling of more than one HCO3- to each Na+. Furthermore, SITS-inhibitable Na(+)-dependent alkalinization exceeds amiloride-inhibitable Na(+)-dependent alkalinization by an order of magnitude, suggesting that the transport capacity of Na(+)-coupled HCO3- transport exceeds that of Na(+)-H+ exchange.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Dengue-1 virus envelope glycoprotein gene expressed in recombinant baculovirus elicits virus-neutralizing antibody in mice and protects them from virus challenge.

In order to test the feasibility of baculovirus (Autographa californica nuclear polyhedrosis virus, AcNPV) expression vectors for making immunogens against dengue-1 (DEN-1) virus, a portion of the envelope (E) glycoprotein gene of DEN-1 virus was cloned and expressed. The recombinant baculovirus contains 107 nucleotides from the 3' terminus of the DEN-1 matrix (M) gene, which encodes a hydrophobic signal peptide and extends through the first 1, 245 nucleotides of E, terminating 243 nucleotides before the 3' terminus of E. When the recombinant virus was grown in Spodoptera frugiperda cells, about 1 mg of E antigen was made per 10(9) cells. Recombinant E antigen reacted with E protein-specific monoclonal antibodies and stimulated production of DEN-1 virus neutralizing antibody in BALB/c mice. Mice immunized with recombinant E antigen or with heat-inactivated DEN-1 virus were protected significantly against lethal DEN-1 virus challenge. A dose/response effect was observed, with increasing amounts of recombinant antigen leading to increased survival. These results demonstrate the utility of baculovirus for producing immunogens against DEN-1 virus.

Animals↗

Inhibition of protein kinase C activation by low concentrations of aluminium.

An extract of rat brain protein kinase C was fully activated at a total calcium concentration 3 x 10(-4) mol/l (Ca2+ 10(-4) mol/l) in the presence of non-limiting concentrations of phosphatidylserine. Available aluminium (ionised plus soluble hydrolysed) at a median value of 2 x 10(-8) mol/l inhibited enzyme activation by 90% at this non-limiting calcium concentration. Literature values for tissue aluminium concentrations suggest that interference with this key intracellular regulatory protein might occur in vivo.

Aluminum↗

Aluminum absorption by rat duodenum: further evidence of energy-dependent uptake.

Duodenums of intact anaesthetised rats have been perfused with saline-bicarbonate buffer (pH 8.5), containing AlCl3, across a range of concentrations 0-300 microM. A linear relation between rate of uptake and perfusate Al concentration was found. The clearance of Al from the lumen was also studied with the mean perfusate concentration of 60 microM, in the presence of certain inhibitors. The clearance was significantly reduced by NaCN(1 mM) or dinitrophenol (0.1 mM) in the perfusate, and by prior perfusion with vanadate (10 microM) in saline. No effect was observed after colchicine (25 micrograms) was administered i.v. 3 h before perfusion, but vincristine (10 micrograms) given similarly reduced clearance significantly. Verapamil (25 micrograms/ml) in the perfusate caused a small, just significant, reduction in clearance. These data further characterise duodenal absorption of Al under physiological conditions and suggest that energy-dependent transport plays an important role in its uptake. Calcium channels may provide an additional entry site.

Aluminum↗

Protection of mice against lethal Japanese encephalitis with a recombinant baculovirus vaccine.

Genes coding for the E and NS1 glycoproteins of Japanese encephalitis virus (JEV) were cloned into baculovirus expression vectors. The recombinant baculoviruses obtained were used to infect Spodoptera frugiperda cells. The infected cells were used to immunize C57/B mice, which were then challenged with live JEV. Survival was increased from about 30% in unimmunized mice to 70% in E and polyprotein recipients (P less than 0.005), but was not increased in NS1 recipients despite the development of antibody against NS1 by these mice. Virus neutralizing antibody was demonstrated in 18/20 E glycoprotein recipients and 15/20 polyprotein recipients. The baculovirus expressed E glycoprotein stimulated antibody which was protective and neutralizing in this system.

Animals↗

Evidence implicating MHC genes in the immunological nonresponsiveness to the Plasmodium falciparum CS protein.

The circumsporozoite (CS) protein is a major candidate vaccine antigen for the sporozoite stage of malaria. Both cytotoxic T cells (CTL) and antibody specific for the CS protein are thought to be important in protection. By examining the immune response in mice and humans we have shown that genes mapping to the major histocompatibility complex (MHC) are important for immune responsiveness. F1 mice between high antibody responders and low antibody responders are high antibody responders, suggesting that in this model immune suppressor genes do not control the immune response. Using synthetic peptides to map epitopes for CTL and helper T cells (which are important for the antibody response) we have shown that the T-cell epitopes are located in the polymorphic region of the protein, and we hypothesize that T cells have indeed selected the variation observed in the CS protein. The success of subunit vaccines will depend on the pattern of variation in different geographical locations, the ability to construct multivalent vaccines containing different variant epitopes from this protein, and on the existence of other sporozoite and liver-stage proteins involved in protection.

Animals↗