Showing papers by "University of Alabama at Birmingham published in 1987"

Immunoglobulin A (IgA): molecular and cellular interactions involved in IgA biosynthesis and immune response.

Abstract: Publisher Summary This chapter discusses the impact of new information concerning IgA physiology on the immune system. IgA should not be considered only as an isotype providing specific humoral protection of mucosal surfaces but as an integral component of the entire immune system. An unusual structural feature of human IgA is the heterogeneity of the molecular forms with characteristic distribution in various body fluids. Though most IgA in serum displays a typical four-polypeptide chain structure of the basic molecule with two Q and two light (L) chains, external secretions contain dimeric and tetrameric, disulfide-linked molecules associated with additional polypeptides-J (joining) chain and secretory component (SC). IgA-producing plasma cells are distributed in various lymphoid and nonlymphoid tissues and are particularly preponderant in the lamina propria of the gut; in salivary, lacrimal, and lactating mammary glands; and in the human bone marrow. IgA occurs in different body fluids in predominantly polymeric or monomeric (plasma, cerebrospinal fluid) forms with a characteristic distribution of IgAl and IgAz molecules. Under normal conditions, an absolute majority of IgA-containing cells in secretory glands and tissues also contain J chain whereas such cells in, for example, normal bone marrow does not. Staining with fluorochrome-labeled anti-J chain is enhanced by the pretreatment of alcohol-fixed tissue sections with acid urea, which leads to the exposure of masked antigenic determinants of intracellular J chain. Specialized lymphoid tissues associated with mucosal surfaces play an essential role in the induction and regulation of generalized immune responses in external secretions.

The common mucosal immune system and current strategies for induction of immune responses in external secretions

Abstract: The selective induction of antibodies in external secretions is desirable for the prevention of various systemic as well as predominantly mucosa-restricted infections. An enormous surface area of mucosal membranes is protected primarily by antibodies that belong, in many species, to the IgA isotype. Such antibodies are produced locally by large numbers of IgA-containing plasma cells distributed in subepithelial spaces of mucosal membranes and in the stroma of secretory glands. In humans and in some animal species, plasma-derived IgA antibodies do not enter external secretions in significant quantities and systemically administered preformed IgA antibodies would be of little use for passive immunization. Systemic administration of microbial antigens may boost an effective S-IgA immune response only in a situation whereby an immunized individual had previously encountered the same antigen by the mucosal route. Local injection of antigen in the vicinity of secretory glands is usually accompanied by an undesirable concomitant systemic response and frequently requires the addition of adjuvants that are unacceptable for administration in humans. Immunization routes that involve ingestion or possibly inhalation of antigens lead to the induction of not only local but also generalized immune responses manifested by the parallel appearance of S-Iga antibodies to ingested or inhaled antigens in secretions of glands distant from the site of immunization. Based on extensive studies in animal models as well as in humans, convincing evidence is available that antigen-sensitized and IgA-committed precursors of plasma cells from GALT are disseminated to the gut, other mucosa-associated tissues, and exocrine glands. However, due to the limited absorption of desired antigens from the gut lumen of orally immunized individuals, repeated large doses of antigens are required for an effective S-IgA response. Novel antigen delivery systems for the stimulation of such responses are currently being examined in several laboratories. Live attenuated or genetically manipulated bacteria expressing other microbial antigens have also been used for selective colonization of gut-associated lymphoid tissues. Unique antigen packaging and the use of adjuvants suitable for oral administration hold promise for an efficient antigen delivery to critical tissues in the intestine and deserve extensive exploration. The oral immunization route appears to have many advantages over systemic immunization.(ABSTRACT TRUNCATED AT 400 WORDS)

Clinical Pharmacokinetics of 5-Fluorouracil and Its Metabolites in Plasma, Urine, and Bile

Abstract: Kinetics of 5-fluorouracil (FUra) and FUra metabolites in plasma and urine were investigated in 10 cancer patients following i.v. bolus administration of 500 mg/m2 FUra with 600 microCi of [6-3H]FUra. Biliary excretion was examined in two patients with external biliary catheters. Quantitation of unchanged drug and metabolites was assessed by a highly specific high-performance liquid chromatographic method. FUra plasma levels declined rapidly with an apparent elimination half-life of 12.9 +/- 7.3 min. Dihydrofluorouracil was detected within 5 min in most patients, demonstrating rapid catabolism and reached maximum peak levels of 23.7 +/- 9.9 microM at approximately 60 min. The apparent elimination half-life of dihydrofluorouracil (61.9 +/- 39.0 min) was consistently greater than that of the unchanged drug. The apparent elimination half-lives of the subsequent metabolites alpha-fluoro-beta-ureidopropionic acid and alpha-fluoro-beta-alanine were prolonged with values of 238.9 +/- 175.4 min and 1976 +/- 358 min, respectively. Approximately 60-90% of the administered dose was excreted in urine within 24 h, primarily as alpha-fluoro-beta-alanine. Biliary excretion accounted for 2-3% of total administered radioactivity. The major fraction of this radioactivity eluted on high-performance liquid chromatography as a previously unrecognized FUra metabolite. Analysis of its structure is currently ongoing in our laboratory. In conclusion, this study provides the first comprehensive analysis of the formation and excretion of FUra metabolites in plasma, urine, and bile following i.v. bolus administration of FUra in humans.

Phosphorylation determines two distinct species of Tau in the central nervous system.

Abstract: The monoclonal antibody, Tau-1, which had previously been used to localize tau to the axonal compartment in brain has been reutilized for light and electron microscopic immunohistochemistry following phosphatase treatment of tissue. We report here that a significant quantity of tau in the central nervous system is phosphorylated in situ at or near the Tau-1 epitope, preventing the binding of the Tau-1 antibody. Upon removal of this/these phosphate group(s), however, Tau-1 was observed in the somatodendritic compartment of neurons as well as in axons. Furthermore, intense staining was also observed in astrocytes and in perineuronal glial cells. This immunoreactivity was present along the lengths of microtubules and on ribosomes (polysomes). Treatment of immunoblots of extracts of whole cerebral cortex with phosphatase confirmed the immunohistochemical results in that a 50-65% increase in Tau-1 binding to the tau region of the blot was noted. Moreover, a novel monoclonal antibody, Tau-2, was also used in these experiments. This antibody binds only to tau and localizes along microtubules in axons, somata, dendrites, and astrocytes and on ribosomes (polysomes) without phosphatase pretreatment.

Interactions between the neuroendocrine and immune systems: common hormones and receptors.

Abstract: The studies reviewed here support a molecular basis for bidirectional communication between the immune and neuroendocrine systems. The main findings can be summarized as follows: First, cells of the immune system can synthesize biologically active neuroendocrine peptide hormones. Second, immune cells also possess receptors for many of these peptides. Third, these same neuroendocrine hormones can influence immune function; and fourth, lymphokines can influence neuroendocrine tissues. Although recent studies have begun to unravel the biochemistry of bidirectional communication between the immune and neuroendocrine systems, there are still missing parts in this puzzle. Among the important questions that must be resolved are the identification of factors that trigger the synthesis of neuroendocrine hormones by immune cells. Are these events operating similar to or in balance with pituitary cells? Drugs that interfere with either pathway may be useful. Second, it will be of value to understand the factors controlling neuroendocrine hormone receptor expression on immune cells. A better understanding of the spectrum of positive and negative regulatory events for both systems may determine the ultimate behavior of immune and neuroendocrine cells. In addition, since leukocytes can produce hormones and also have receptors for the same hormones (e.g., ACTH and GH), it is possible that these immunocytes may also influence their own function in an autocrine-like fashion. We have postulated that the immune system can serve as a sensory organ for external stimuli that cannot be detected by the nervous system (Blalock 1984). Thus, the immune system recognizes stimuli such as bacteria, viruses or tumors, whereas the nervous system detects classical sensory stimuli. The contribution of extrapituitary sites of hormone production and function may provide new clues to define psychological and/or pathological states in the pathophysiology of infectious diseases and tumors.

Contrast sensitivity, acuity, and the perception of 'real-world' targets.

Abstract: A major assumption underlying the use of contrast sensitivity testing is that it predicts whether a patient has difficulty seeing objects encountered in everyday life. However, there has been no large-scale attempt to examine whether this putative relationship actually exists. We have examined this assumption using a clinic based sample of adults aged 20-77 years. Contrast thresholds were measured for both: (1) gratings of 0.5-22.8 cycles/degree; and (2) real-world targets (faces, road signs, objects). Multiple regression techniques indicated that the best predictors of thresholds for real-world targets were age and middle to low spatial frequencies. Models incorporating these variables accounted for 25-40% of the variance. Although acuity significantly correlated with thresholds for real-world targets, the inclusion of acuity as a predictor variable did not improve the model. These data provide direct evidence that spatial contrast sensitivity can effectively predict how well patients see targets typical of everyday life.

Phosphorylation Fails to Activate Chloride Channels from Cystic Fibrosis Airway Cells

Abstract: Chloride impermeability of epithelial cells can account for many of the experimental and clinical manifestations of cystic fibrosis (CF)1,2. Activation of apical-membrane Cl− channels by cyclic AMP-mediated stimuli is defective in CF airway epithelial cells3,4, despite normal agonist-induced increases in cellular cAMP levels4,5. This defect in Cl− channel regulation has been localized to the apical membrane by exposing the cytoplasmic surface of excised membrane patches to the catalytic subunit (C subunit) of cAMP-dependent protein kinase and ATP. In membranes from normal cells, C-subunit activated Cl− channels with properties identical to those stimulated by cAMP-dependent agonists during cell-attached recording. Activation by the C subunit was not observed in CF membranes, but the presence of Cl− channels was verified by voltage-induced activation. The failure of the C subunit to activate the Cl− channels of CF membranes indicates that the block in their cAMP-mediated activation lies distal to induction of cAMP-dependent protein kinase activity and focuses our attention on the Cl− channel and its membrane-associated regulatory proteins as the probable site of the CF defect.

Psychological, social, and health consequences of caring for a relative with senile dementia.

Abstract: While providing home care for a family member with senile dementia is clearly extremely stressful, there has been little controlled research assessing the specific effects of this stress on caregiver psychological, social, and health functioning. To address this question, 44 primary caregivers of senile dementia patients and 44 matched controls completed a series of questionnaires and interview assessments. Caregivers reported significantly higher levels of depression and negative affect toward their relatives, and lower overall life satisfaction than controls. Caregivers also had significant impairment of their social activities, including visits with friends, vacations, and church attendance when compared with controls. Caregivers expressed less satisfaction with their social networks than did controls, but the groups did not differ in objective size of social network or number of network contacts. Caregivers reported poorer health, more prescription medication use, and higher utilization of health care than controls. Results clearly indicate the serious and wide-ranging effects of the stress of caregiving, and reinforce the importance of providing comprehensive services for caregiving families.

Organization of central adrenergic pathways : I. Relationships of ventrolateral medullary projections to the hypothalamus and spinal cord

Abstract: We studied the organization of projections from the C1 adrenergic and A1 noradrenergic cell groups in the ventrolateral medulla (VLM) to the hypothalamus and the spinal cord by using a combination of retrograde transport of fluorescent tracers and immunocytochemistry. Three issues were addressed. Neurons in the VLM that stain immunohistochemically for phenylethanolamine N-methyltransferase (PNMT) have been assumed to be adrenergic. However, the presence of PNMT-immunoreactive neurons in the hypothalamus that do not stain for tyrosine hydroxylase (TH) prompted us to re-evaluate the VLM by an elution-restaining immunohistochemical procedure. We confirmed that nearly all of the rostral medullary PNMT-immunoreactive neurons also stained for TH. By contrast, in the caudal medulla, very few TH-positive neurons stained for PNMT. Neurons of the C1 group in the rostral VLM project both to the thoracic spinal cord and to the hypothalamus. To determine whether individual C1 neurons send collaterals to the hypothalamus and spinal cord, we injected different-colored fluorescent dyes (diamidino yellow or fast blue) into the thoracic spinal gray matter and either the median preoptic (MnPO) or paraventricular (PVH) nuclei of the hypothalamus. Very few double-labeled neurons were found in the VLM, indicating that hypothalamic and spinal cord projections arise from almost completely independent populations of cells. Approximately half of the neurons projecting to the spinal cord from rostral VLM were not immunoreactive for TH or PNMT, indicating that a substantial part of this projection is noncatecholaminergic. The MnPO and the PVH both receive extensive catecholaminergic inputs from the VLM. We also used fluorescent retrograde tracers to determine whether individual VLM neurons send collaterals to both hypothalamic sites. Approximately 20% of neurons projecting to the MnPO in the rostral two thirds of the VLM also sent collaterials to the PVH, nearly all of these neurons being TH-positive. The collateralization of the VLM catecholaminergic projection to the hypothalamus may provide an anatomical substrate for integration of fore-brain participation in cardiovascular regulation. In contrast, the adrenergic projection from the VLM to the intermediolateral column of the spinal cord arises from a separate population of neurons.

The concept of a critical oxygen delivery.

Abstract: In healthy tissues, decreases in oxygen delivery (QO2 = cardiac output X arterial O2 content) do not lower oxygen consumption (VO2) because tissue O2 extraction increases proportionately When delivery is reduced below a critical threshold, VO2 falls because tissue extraction exceeds a critical threshold, and cannot compensate for the reduction in delivery In the adult respiratory distress syndrome and perhaps in septicemia, tissue extraction capacity is impaired, leading to O2 supply dependency despite normal or increased overall delivery This pathologic supply dependency could be caused by a loss in autoregulatory capacity, by disrupted blood flow distribution secondary to peripheral microembolization, or to other factors interfering with efficient tissue distribution of QO2 with respect to VO2 Alternatively, the increased VO2 may be consumed in biochemical pathways not associated with ATP production, or in the production of oxygen radicals or hydrogen peroxide To the extent this abnormal dependence of VO2 on QO2 reflects tissue hypoxia, clinical interventions which decrease systemic delivery should be evaluated with regard to possible deleterious effects on organ system function

Enhancement of neutrophil superoxide production by preincubation with recombinant human tumor necrosis factor.

Abstract: Tumor necrosis factor (TNF) is a 17,000-Da protein which is produced by mononuclear cells upon exposure to endotoxin. Increases in adherence, phagocytosis, hydrogen peroxide release, and lysozyme secretion have been demonstrated after prolonged incubation of human neutrophils with TNF. In this study, the ability of highly purified recombinant human TNF to modulate neutrophil responses to soluble stimuli was evaluated. Tumor necrosis factor alone (0.1 to 10,000 units/ml) failed to induce neutrophil superoxide anion (O2-) production, granule release, or aggregation when incubated for up to 25 min at 37 degrees C. TNF did, however, stimulate a significant time-, dose-, and temperature-dependent increase in neutrophil F-actin content. Although exposure of neutrophils to TNF alone caused no superoxide anion production, it enhanced the O2- production in response to the chemotactic peptide, f-methionyl-leucyl-phenylalanine (FMLP) or the tumor promotor, phorbol myristate acetate, by as much as 278%. The enhancement was time-, dose-, and temperature-dependent and was due to a more rapid initial rate of O2- production. The TNF enhancement of FMLP-induced O2- production was blocked when an anti-TNF monoclonal antibody 241-1H11, is present during the preincubation period. TNF preincubation also enhanced FMLP-induced lysozyme release, but had no effect on aggregation and actin polymerization by FMLP. The kinetics of NADPH oxidase activation by arachidonic acid was unaltered by TNF. These results indicate that brief exposures to recombinant human TNF are able to enhance or prime the neutrophil oxidative burst in response to a second stimulus.

Fetal Methylmercury Poisoning: Relationship Between Concentration in Single Strands of Maternal Hair and Child Effects

Abstract: • Pregnant women consumed bread that was prepared from methylmercurytreated wheat. Single strands of maternal head hair were analyzed by x-ray fluorescence spectrometry. The index of fetal exposure was the maximum hair mercury concentration during gestation. Effects were measured by the frequency of psychomotor retardation, seizures, and neurological signs in the children. A dose-response relationship was demonstrated for fetal effects of methylmercury. Analysis of single hair strands provides a better index of acute or subacute fetal exposure than analysis of bundles of hair; the duration and degree of exposure are more accurately defined. A sex difference in response is discussed.

A Critical Period for Learning to Pronounce Foreign Languages

Abstract: This article discusses the Critical Period Hypothesis (CPH) as it relates to the naturalistic acquisition of foreign-language (L2) pronunciation by adults and children. An examination of the existing empirical and theoretical literature leads to the conclusion that there is no conclusive support for the existence of a critical period for human speech learning, and that assuming a critical period does exist may inhibit the search for testable hypotheses concerning the basis for observed adult-child differences in L2 pronunciation. These conclusions are based on the existence of direct coullter-evidence, as well as on the observation that apparent adult-child performance differences may arise from many different confounding factors other than adult-child differences in neurological maturation or organization that cannot be adequately controlled in behavioral research.

Left-handed DNA in vivo

Abstract: Left-handed DNA is shown to exist and elicit a biological response in Escherichia coli. A plasmid encoding the gene for a temperature-sensitive Eco RI methylase (MEco RI) was cotransformed with different plasmids containing inserts that had varying capacities to form left-handed helices or cruciforms with a target Eco RI site in the center or at the ends of the inserts. Inhibition of methylation in vivo was found for the stable inserts with the longest left-handed (presumably Z) helices. In vitro methylation with the purified MEco RI agreed with the results in vivo. Supercoil-induced changes in the structure of the primary helix in vitro provided confirmation that left-handed helices were responsible for this behavior. The presence in vivo of left-handed inserts elicits specific deletions and plasmid incompatibilities in certain instances.

1,25-Dihydroxyvitamin D3 regulates the biosynthesis of osteopontin, a bone-derived cell attachment protein, in clonal osteoblast-like osteosarcoma cells.

Abstract: We investigated the effects of 1,25-dihydroxyvitamin D3 on the synthesis of osteopontin, a phosphorylated cell attachment glycoprotein, in ROS 17/2.8 cells, a clonal osteoblast-like rat osteosarcoma cell line. We observed a dose dependent increase in uptake of [32PO4] into osteopontin secreted into the medium. An increased incorporation of [35S]-methionine into secreted osteopontin suggested the effect was that of increased protein biosynthesis. Using a radioimmunoassay we demonstrated a dose dependent increase in the amount of secreted osteopontin, an increase which could be blocked by Actinomycin D, in response to 1,25-dihydroxyvitamin D3. These results suggest that the hormonal form of vitamin D regulates the biosynthesis of osteopontin, possibly at the level of transcription.

Anatomical observations on the acromioclavicular joint and supporting ligaments

Abstract: An anatomical study of the acromioclavicular (AC) joint and its supporting ligaments was performed using both macroscopic and microscopic methods. The project used 63 cadaver shoulders of unknown ages. Fifty- three joints were used for macroscopic and 10 for microscopic study. The data consisted of 1) micrometer measurements of the dimensions of the extrinsic and intrinsic ligaments of the AC joint; 2) measurement and description of the intraarticular meniscus and the su perior and inferior capsular ligaments of the AC joint; and 3) the anatomical course and relationship of the coracoacromial (CA) ligament to the supporting liga ments of the AC joint and a description of its insertion on the acromion process.The following observations were made: The cor acoclavicular ligament, especially the trapezoid liga ment, provides significant soft tissue restraints to up ward displacement of the clavicle. A complete AC joint disc was found in only one, meniscoid discs in 25, remnants of discs in 16, and no discs in...