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Biomedical subjects

J James

Publications and source records attributed to J James.

At least 109 records · Page 6Linked to original sources
PubMed1990

Use of plasma with high levels of ionised calcium in the production of model scale coagulation factor concentrates.

We have attempted to exploit the Ca2(+)-dependent stability of factor VIII in producing factor VIII concentrates of higher yield. Plasma levels of ionised calcium were increased in two ways: (a) whole blood collection into half-strength citrate CPD anticoagulant, leading to free Ca2+ levels of ca 120 microM and (b) apheresis collection of plasma which was then recalcified to free Ca2+ levels of ca 300 microM under heparin cover. Coagulation factor concentrates were prepared using model versions of our industrial scale manufacturing methods. Factor VIII yield was increased through low citrate collection. This did not compromise factor IX yield or thrombogenic potential. Use of recalcified heparinised plasma did not lead to any improvement in factor VIII yield and resulted in a marked drop in factor IX recovery, possibly from interference by heparin of factor IX binding in ion-exchange chromatography. The benefits accruable through the use of half-strength citrate CPD anticoagulant support the continued evaluation of this preservative in large scale blood collection and fractionation. The deleterious effects of heparin in charge-mediated plasma fractionations may pose serious difficulties in harvesting vitamin K dependent factors.

Anticoagulants↗
PubMed1990

Elastic and collagenous fibers in the temporomandibular joint capsule of the rabbit and their functional relevance.

This study deals with the form and function of the temporomandibular joint capsule and compares the histological structure of the capsule and kinetics of condylar and disc movement in rabbit and man. The morphology of the capsule of the jaw joint of the rabbit was studied by means of histological sections of isolated capsules, celloidin sections of decalcified heads, and cryostat sections containing the joint, fixed in jaw open and closed positions. General staining methods and selective methods for elastin and collagen were used. The volume density of the elastic fibers was determined morphometrically. The capsule consists of a portion connecting disc and skull and a portion connecting disc and mandible. The former portion has a fibrous outer layer of dense connective tissue. It contains numerous bundles of specifically oriented collagen but hardly any elastic fibers. Few irregularly arranged elastic fibers are present in the lateral and medial areas of the capsule. In contrast, the portion connecting disc and condylar process contains not only oriented collagen but also large amounts of elastic fibers. Anteriorly, there is a thin elastic band that is stretched greatly at jaw opening. Posteriorly, the capsule takes the appearance of a large fibro-elastic pad that covers the posterior extension of the condyle. This part is somewhat stretched when the jaw is closed. The findings correlate well with the results of jaw movement studies, showing a remarkable capability for condylar protrusion and retrusion. In contrast to the situation in humans, this movement occurs mainly between condyle and disc. It is made possible by the rather tight connections between skull and disc and flexible ones between disc and condyle.

Animals↗
PubMed1990

Immunocytochemical determination of ploidy class-dependent bromodeoxyuridine incorporation in rat liver parenchymal cells after partial hepatectomy.

Immunocytochemistry of bromodeoxyuridine (BrdU) incorporated in DNA was performed on cryostat sections of rat liver and on isolated hepatocytes after partial hepatectomy using a two-step labeling technique. The method enabled the detection of S-phase nuclei in both tissue preparations. Quantification of the number of labeled nuclei in sections showed that the number of nuclei in S-phase increased from 0.3% in control liver to about 36% at 24 h after partial hepatectomy. The detection of BrdU in isolated hepatocytes showed the same labeling index of binuclear diploid, mononuclear tetraploid and binuclear tetraploid cells. A special role for mononuclear diploid cells in proliferation did not seem to occur.

Animals↗
PubMed1990

Silica-associated systemic sclerosis is clinically, serologically and immunologically indistinguishable from idiopathic systemic sclerosis.

To determine whether the clinical, immunological and serological features of patients with silica-associated systemic sclerosis are different from patients with the 'idiopathic' form of systemic sclerosis (SS) we studied 22 underground coal miners who were exposed to silica dust (SD), 30 mine workers who later developed silicosis (S) and 17 mine workers exposed to silica dust who subsequently developed a systemic sclerosis-like disease (SA-SS). The patients with SA-SS had features clinically indistinguishable from individual patients with SS. They all had Raynaud's phenomenon, 14 had cutaneous sclerosis identical to that seen in acrosclerosis and three had a generalized cutaneous sclerosis. Sixteen patients had bibasilar pulmonary fibrosis, 10 had necrosis of the fingertip pulps, nine had oesophageal involvement and only one patient had renal involvement. Antinuclear antibodies and circulating immune complexes were detected in three and eight patients with SD, 14 and five patients with S and in 16 and nine patients with SA-SS, respectively. Anti-Scl-70 antibody was detected in eight of the 17 patients with SA-SS. Evidence for in vivo endothelial cell damage, as determined by elevated levels of von Willebrand factor, was found in nine patients with SD, 14 patients with S and in 10 patients with SA-SS. Following incubation of the patient's serum with confluent cultures of human umbilical vein endothelial cells there was only a significant reduction in calcium ionophore-induced release of prostacyclin with the serum from SA-SS patients compared to that with control serum (NC). The mean +/- SEM release of 6-keto-PGF1 alpha (the stable metabolite of prostacyclin expressed as ng/10(4) cells) decreased from 2.90 +/- 0.27 to 2.01 +/- 0.33 (SD), 3.34 +/- 0.42 to 1.76 +/- 0.31 (S), 1.98 +/- 0.12 to 0.64 +/- 0.07 (SA-SS) and 2.28 +/- 0.33 to 1.36 +/- 0.21 (NC) with 1 and 20% serum, respectively. This study demonstrates that immune complex and antinuclear antibody formation and in vivo endothelial cell damage occurs following occupational exposure to silica. The patients who subsequently develop a systemic sclerosis-like disease have clinical, immunological and serological features which are indistinguishable from the idiopathic form of the disease although as a group the SA-SS patients have a higher prevalence of pulmonary involvement and the anti-Scl-70 antibody.

6-Ketoprostaglandin F1 alpha↗
PubMed1990

Sirius red histophotometry and spectrophotometry of sections in the assessment of the collagen content of liver tissue and its application in growing rat liver.

By means of staining with Sirius Red F3BA in a saturated picric acid solution, the collagen contents of rat livers with varying degrees of fibrosis have been measured quantitatively in fixed and sectioned material, using both histophotometry in situ and extraction of bound dye with colorimetric analysis. These findings have been correlated with chemical assays of the hydroxyproline content in homogenates from the same livers. It appears that a highly significant correlation exists between both section-based analysis methods and the hydroxyproline content, the Spearman-Rank correlation coefficients being virtually identical. For analysis of collagen accumulation in rat liver, both section-based methods seem to be useful and reliable, the extraction method giving the quickest results for large-scale screening, and the histophotometric method being more appropriate to take readings in selected areas. With human liver material, indications have been obtained for the existence of large sampling errors due to inhomogeneous distribution of collagen deposits. Using the extraction method, no significant changes could be observed in the volume density of collagen during postnatal growth from 1 week to 21 months in rat liver: only on the third day after birth was a higher value of collagen/total protein obtained, possibly due to a higher water content of the hepatocytes. Partial hepatectomy was found to have no influence at all on the collagen content of rat liver during the period of restorative growth or after it.

Animals↗
PubMed1990

Quantitative changes in acid phosphatase, alkaline phosphatase and 5'-nucleotidase activity in rat liver after experimentally induced cholestasis.

Acid phosphatase, alkaline phosphatase and 5'-nucleotidase activities were analyzed cytophotometrically in cryostat sections of rat liver up to 8 weeks after ligation and transsection of the common bile duct. Ligation resulted in cholestasis and induced alterations in both localization and activity of the enzyme investigated. The cellular distribution but not the activity of acid phosphatase changed in liver parenchyma. In control liver, the final reaction product was localized as discrete granules in the bile canalicular region of hepatocytes. The final reaction product was precipitated more diffusely within the cytoplasm after induction of cholestasis, most probably due to increased fragility of lysosomal membranes. In control liver, alkaline phosphatase activity was low and localized in the bile canalicular plasma membranes only. The total parenchymal activity increased threefold after the induction of cholestasis and is considered to be a compensatory mechanism in order to enhance the excretion of bile salts from hepatocytes. 5'-Nucleotidase was present at the bile canalicular and sinusoidal surfaces of plasma membranes of hepatocytes in control liver; total activity in pericentral areas was significantly higher than in periportal areas. Induction of cholestasis resulted in higher total activity and redistribution of the activity over all three surfaces of the plasma membranes, whereas heterogeneity over the different zones of the acinus disappeared. The appearance of the enzyme at lateral plasma membranes is suggested to be related to the formation of new sites for bile salt transport out of the hepatocytes. With respect to all three enzymes studied, alterations of liver parenchymal cells due to a disturbed bile transport were already established during the first week of cholestasis.

5'-Nucleotidase↗
PubMed1989

Preventing iron deficiency in preschool children by implementing an educational and screening programme in an inner city practice.

OBJECTIVE: To assess the feasibility and acceptability of screening young children for iron deficiency in a deprived inner city practice and to assess the effects of a programme of dietary education. DESIGN: Prospective study of children in general practice, comparison with historical controls. SETTING: A deprived inner city practice. PATIENTS: 127 Children aged 13-24 months. Findings were compared with those in 110 children of the same age studied previously. INTERVENTIONS: All mothers received dietary education antenatally and in the first year after giving birth. Screening for iron deficiency (defined as mean cell volume less than 75 fl and haemoglobin concentration less than 105 g/l) and haemoglobinopathy (when appropriate) was offered for all children attending for immunisation against measles, mumps, and rubella over 12 months; capillary blood samples were taken after immunisation. MAIN OUTCOME MEASURES: Uptake of the screening programme expressed as the percentage of all children eligible for immunisation who were screened, and the effectiveness of the dietary education as shown by the prevalence of iron deficiency in the two groups. RESULTS: Altogether, 122 of the 127 (96%) children who attended for immunisation had their haemoglobin concentration and mean cell volume measured; 90% of all children aged 13-24 months in the practice were screened. Dietary education, clinical procedures, and counselling were incorporated successfully into the clinic's work. Ten children (8%) were iron deficient, all of whom responded to iron supplements, and eight had a haemoglobinopathy trait. In the previous study 110 children (70%) had been screened and 28 children (25%) had been iron deficient. The two groups were similar in terms of sex, social class, and ethnic group. CONCLUSIONS: Screening young children for iron deficiency, sickle cell disease, and thalassaemia when they attended for immunisation was acceptable and successful in a socially deprived inner city practice. Dietary education may have accounted for some of the reduction in the prevalence of iron deficiency that occurred over the two years.

Anemia, Hypochromic↗
PubMed1989

Prevention of transfusion-acquired cytomegalovirus infection in infants by blood filtration to remove leucocytes. Neonatal Cytomegalovirus Infection Study Group.

A multicentre, controlled trial was carried out to determine whether removal of leucocytes from blood by means of 'Imugard IG500' (Terumo) filters would prevent transfusion-acquired cytomegalovirus (CMV) infection in newborn infants. 72 infants whose mothers were seronegative and who received some seropositive blood were followed for 6 months for evidence of CMV infection. There were no significant differences between the groups who received filtered and unfiltered blood in median gestation, birthweight, or amount of seropositive blood received (median volume 32.5 ml and 34.5 ml, respectively). 9 (21%) of the 42 infants who received unfiltered blood and none of the 30 who received filtered blood were infected with CMV. All infected infants weighed less than 1500 g at birth; they represented 31% of very low birthweight (VLBW) infants at risk of CMV infection. None of 24 VLBW infants who received filtered seropositive blood was infected. 1 infected infant died and 5 had clinical features consistent with CMV infection. The results show that transfusion-acquired CMV infection is preventable by filtration of blood through a leucocyte filter. This method has advantages over other methods of removing leucocytes or the use of only seronegative blood for newborn infants.

Antibodies, Viral↗
PubMed1989

Acquisition of the classically conditioned eyeblink response in humans over the life span.

Human subjects ranging in age from 18 to 85 years underwent classical conditioning of the eyeblink response to a tone conditioned stimulus (CS) and an air-puff unconditioned stimulus (UCS). There was a decline in percentage of conditioned responses with age. This decline was most noticeable in subjects over age 50. These conditioning deficits were not due to age-related changes in sensitivity to the tone CS or the air-puff UCS, nor could the conditioning deficits be attributed to an age-related decline in general cognitive abilities or to changes in spontaneous blink rates. The results are discussed in terms of using the classically conditioned eyeblink in humans in conjunction with the classically conditioned nictitating membrane response in rabbits as a model system for studying the neurobiology of age-related conditioning deficits.

Adolescent↗
PubMed1989

Mechanisms of bacterial opsonization by immune globulin intravenous: correlation of complement consumption with opsonic activity and protective efficacy.

To study the mechanism of bacterial opsonization by immune globulin intravenous (IGIV) complement consumption and polymorphonuclear leukocyte (PMNL) membrane receptor (FcRlo, CR1, and CR3)-mediated phagocytosis of Staphylococcus epidermidis, Klebsiella pneumoniae, and groups A and B streptococci were examined. IGIV alone did not consume complement and showed no opsonic activity by itself for these organisms. When these bacteria were preopsonized in IGIV, significant amounts of complement were consumed (44%-94%) and the uptake and killing of bacteria occurred. The in vitro opsonic activity of IGIV for these organisms was significantly correlated with the amount of complement consumed by the IGIV-opsonized bacteria (r = .85, P less than .05). The in vivo protective efficacy of IGIV also appeared to be directly associated with its ability to activate and consume complement (r = 1.0, P less than .001). Antibodies to FcRlo (Leu 11) markedly inhibited phagocytosis of bacteria opsonized in IGIV but not that of bacteria opsonized in specific IgM. Both CR1 and CR3 receptors on PMNLs were involved in uptake, but the contribution of each is different with different organisms.

Animals↗
PubMed1989

Microtitre plate measurement of platelet response to hypotonic stress.

The conventional method of assessing the platelet response to hypotonic stress (HSR) was adapted to allow microtitre plate technology to be used. After water is added to a platelet suspension two sequential readings are taken at 414 nM on a vertical microplate reader. The difference between the second (three minutes) and the first (one minute) was defined as the HSR. This method allowed the relation between platelet concentrate pH and viability to be confirmed, and an HSR value for use in quality control was established. The method correlated well with the conventional technique and permitted measurement of undiluted samples as well as of products with a high free haemoglobin concentration.

Blood Platelets↗
PubMed1989

Application of keratin immunocytochemistry and sirius red staining in evaluating intrahepatic changes with acute extrahepatic cholestasis due to hepatic duct carcinoma.

A series of 10 cases of biliary obstruction due to primary cholangiocarcinoma has been studied with histological and immunocytochemical means. The total duration of cholestasis (as manifested by jaundice) was between 2 and 11 weeks with variable period of preoperative drainage. Liver biopsy specimens taken during surgery for cholangiocarcinoma were investigated for the presence of ductular proliferation and the development of fibrosis, as demonstrated by Sirius Red F3BA collagen staining. The differentiation of epithelial components was evaluated by AEC-immunostaining with chain-specific monoclonal antibodies specifically directed against human keratins type 7, 18 and 19. Keratin 7, normally occurring only in the ductular system, was expressed in hepatocytes at the periphery of the hepatic lobule (zone I) following about 4 weeks' cholestasis, when an increase of ductular profiles in the enlarged portal areas had become manifest. Such keratin 7 positive cells, however, still retained all morphological aspects of hepatocytes. Keratin 19, normally also restricted to the ductular system in liver, is not expressed by zone I hepatocytes even after longer duration (up to 11 weeks) of cholestasis. It is concluded that the increase in ductular profiles during the first week is mainly due do proliferation of pre-existing ductules, while ductular metaplasia occurs in more chronic cholestasis. Development of fibrosis, not always strictly paralleling the multiplication of ductular profiles in sections through a portal tract, represents an early change, and is clearly apparent after 2 weeks of obstruction.

Acute Disease↗
PubMed1988

Quantitative aspects of the parenchyma-stroma relationship in experimentally induced cholestasis.

In order to quantify changes of the parenchyma/stroma relations in the progression of experimentally induced biliary fibrosis in the rat, localisation of lactate dehydrogenase activity and Sirius Red staining were used as criteria to detect parenchymal cells and collagen fibers, respectively. Blood levels of bilirubin, alkaline phosphatase, anti-thrombin III activity, alpha 2-antiplasmin, factor II and factor X were related to the data obtained by histomorphometric measurements in sections gathered 1, 2, 4 and 6 weeks after the onset of cholestasis in three animals and after 8 weeks in one animal. Histophotometry showed a reduction in volume density of the parenchymal cell mass of 96%, 78%, 76%, 62% and 59% of the control values, respectively. During the same period, the collagen increased 5-fold in 4 weeks time, levelling off afterwards. Newly formed collagen appeared in the portal areas in close association with proliferating ductules, invading with the latter into the parenchymal mass. After 6 weeks, regressive changes were observed in the ductule complexes formed, manifested by a lowering of the epithelium in which extensive apoptotic cell death was observed with the electron microscope. Of the blood parameters analyzed, the clotting factor X showed the best inverse correlation with the Sirius Red readings (rs = -0.84), i.e. the volume density of collagenous fibers.

Animals↗
PubMed1988

Mineral accretion in the human fetus.

Bone mineral content of the mid forearm was measured by photon absorptiometry in 73 white singletons (36 boys, 37 girls) born between 18 and 43 weeks' gestation. Results obtained within two weeks of birth for liveborn infants were used to establish bone mineral deposition curves approximating normal in utero development. Results for stillborn infants indicated a retardation of bone mineralisation relative to liveborn infants. This has important implications concerning previous estimates of daily calcium needs for preterm infants. Eleven liveborn Asian singletons and nine pairs of white twins were examined similarly. With respect to bone mineral content, weight, and crown-heel length, Asians with gestational ages over 35 weeks and twins were closely comparable with their white singleton peers. Asians under 35 weeks tended to be smaller than white singletons of comparable age.

Asia↗