Transcriptional regulation of cellular retinaldehyde-binding protein in the retinal pigment epithelium. A role for the photoreceptor consensus element

Cellular retinaldehyde-binding protein (CRALBP) is abundantly expressed in the retinal pigment epithelium (RPE) and Muller cells of the retina, where it is thought to function in retinoid metabolism and visual pigment regeneration. Mutations in human CRALBP that destroy retinoid binding have been linked to autosomal recessive retinitis pigmentosa. To identify the DNA elements that regulate expression of the human CRALBP gene in the RPE, transient transfection studies were carried out with three CRALBP-expressing human RPE cell culture systems. The regions from -2089 to -1539 base pairs and from -243 to +80 base pairs demonstrated positive regulatory activity. Similar activity was not observed with cultured human breast, liver, or skin cells. Since sequence analysis of the -243 to +80 region identified the presence of two photoreceptor consensus element-1 (PCE-1) sites, elements that have been implicated in photoreceptor gene regulation, the role of these sequences in RPE expression was examined. Mutation of either PCE-1 site significantly reduced reporter activity, and mutation or deletion of both sites dramatically reduced activity. Electrophoretic mobility shift analysis with RPE nuclear extracts revealed two complexes that required intact PCE-1 sites. These studies also identified two identical sequences (GCAGGA) flanking PCE-1, termed the binding CRALBP element (BCE), that are also important for complex formation. Southwestern analysis with PCE-1/BCEcontaining probes identified species with apparent masses near 90-100 and 31 kDa. These results begin to identify the regulatory regions required for RPE expression of CRALBP and suggest that PCE-1-binding factor(s) may play a role in regulating RPE as well as photoreceptor gene expression.     

The prevention of cancer

The purpose of this review is to summarize recent functional and structural findings regarding non-collagenous matrix proteins in bone and teeth, to compare gene locations for bone and tooth matrix proteins with loci for hereditary skeletal diseases, and to present several provocative hypotheses which integrate this new information into a physiological context. Hypothesis I proposes that the molecular composition of rapidly deposited and mineralized woven bone, as well as the responsiveness of cells synthesizing woven bone to stimuli, is different from that for more slowly synthesized lamellar bone, implying the existence of distinctive osteogenic mechanisms. This review of recent research strongly supports this proposal. Briefly, the protein composition of woven bone matrix is enriched in acidic phosphoproteins BAG-75 and BSP, which are not expressed in lamellar bone, which is itself enriched in osteocalcin. De novo deposition and mineralization of woven bone occurs faster than in lamellar bone by means of a matrix-vesicle-assisted mechanism. Deposition of woven bone occurs at sites experiencing biomechanical strains higher than those experienced by lamellar bone. In addition, woven bone in metaphyseal regions is more susceptible to osteoclastic resorption after space flight, ovariectomy, and loss of weightbearing than is lamellar bone. Finally, osteoprogenitor cells responsive to parathyroid hormone reside in the metaphyseal region of long bones. Taken together, these findings suggest that Hypothesis I represents a useful paradigm for future studies. Specific functions mediated by most individual bone and tooth matrix proteins remain uncertain. A review of current literature suggests that the functionality of skeletal matrix proteins is expressed through specific binding sites composed of particular species-conserved structural motifs (Hypothesis 2). Examples include the previously recognized Asp-Ser-Ser motif of dentin phosphophoryns and the gamma-carboxyglutamic acid motif of matrix GLA protein and osteocalcin. A new polyacidic amino acid motif composed of consecutive Asp and Glu residues (n > 7) was defined in extracellular matrix components osteopontin, bone sialoprotein, and bone acidic glycoprotein-75 on the basis of strong functional analogies with similar polyacidic stretches in divalent metal storage proteins of the endoplasmic reticulum and sarcoplasmic reticulum. These structural motifs represent prime targets for future structure-function studies in vivo and in vitro.     

Relationship between lead mining and blood lead levels in children

The authors studied blood lead levels of 226 randomly selected children, aged 6-92 mo, who lived in either a lead-mining area or a nonmining area, and 69 controls. The authors sought to determine to what extent mining activities contributed to blood lead levels in the children. The mean blood lead levels in the study and control groups were 6.52 microg/dl and 3.43 microg/dl, respectively. The corresponding proportions of children with elevated blood lead levels were 17% and 3%. Soil and dust lead levels were up to 10 times higher in the study than the control group. Elevated blood lead levels appeared to result from exposure to both lead-mining waste and lead-based paint. Mining waste was the cause of the higher prevalence of elevated blood lead levels in these children.     

The deamination of L-arginine and the synthesis of active nitrogen compounds in Yersinia pestis

The investigations of authors concerning infants and their families were generalized on the basis of the observations of infants and their parents with some psychological problems in families. The principles of psychotherapeutic work in above-mentioned groups were formulated. Besides the methods of the diagnosis of psychical deviations development in first year infants as well as peculiarities of alterations in mother-baby system and the main forms of psychic disturbances correction in infants were also described.     

Cardiac troponin-I accurately predicts myocardial injury in renal failure

BACKGROUND: Non-specific elevations of creatine kinase isoenzymes (CK-MB) and cardiac troponin-T may be seen in renal failure, confusing the diagnosis of myocardial infarction. Cardiac troponin-I (cTn-I) has been shown to be specific for myocardial damage in several disease states, but has not been prospectively evaluated in the setting of renal failure. METHODS: This prospective case series evaluated 56 patients with acute or chronic renal failure or end-stage renal disease to assess the sensitivity and specificity of cTn-I for detecting myocardial injury in this patient population. During a 6-month period, patients admitted with suspected myocardial injury by history, physical examination, and electrocardiography were evaluated. Cardiac troponin-I (cTn-I) measurements were assessed between 8 and 48 h after admission. Appropriate medical care and further cardiac testing (echocardiography, stress testing, or arteriography) was performed at the discretion of the primary physician. RESULTS: Myocardial injury was diagnosed in 18/56 (32%) patients by positive cTn-I levels, while only 7/56 (13%) patients had evidence of myocardial damage by CK-MB. Twenty-one of 56 (38%) patients had indeterminate CK-MB levels and 53% of these patients demonstrated myocardial ischaemia on follow-up testing. Sixteen patients had negative cardiac studies; all of these patients had negative cTn-I levels, while seven of these 16 (44%) patients had indeterminate CK-MB measurements. All of the patients with positive cTn-I levels had positive cardiac studies. Positive troponin levels were associated with increased in-hospital mortality. Sensitivity and specificity for CK-MB were 44 and 56% respectively, and 94 and 100% for cTn-I. CONCLUSION: These data support the use of cTn-I for diagnosing myocardial injury in patients with renal failure. Elevated cTn-I levels are associated with increased short-term mortality in renal failure patients. The accuracy of cTn-I could potentially limit unnecessary cardiac testing in renal failure patients, while the enhanced sensitivity contributes to risk stratification and aids in diagnosing true myocardial injury in this population susceptible to non-specific elevations in other muscle enzymes.     

Modification of Tefzel film through graft copolymerization

The graft copolymerization of styrene (st) and methacrylonitrile (MAN) onto Tefzel film in aqueous media by the preirradiation method has been studied. In order to follow the effect of preswelling of the backbone polymer, grafting was attempted onto preirradiated Tefzel film and monomer preswollen, preirradiated Tefzel film. Optimum conditions pertaining to maximum percentage of grafting of st and MAN have been evaluated. Grafting onto preswollen, preirradiated Tefzel film displayed better results. The effect of different alcohols of increasing chain length on the percentage of grafting of st and MAN was also studied. Graft copolymerization of st showed an increase, while grafting with MAN exhibited a decrease, in the percentage of grafting in the presence of alcohols as compared to that obtained in the aqueous medium. Characterization of the graft copolymers was made by IR and thermogravimetric studies. Tefzel‐graft‐polystyrene showed improved thermal stability while the MAN grafted onto preswollen, preirradiated Tefzel film produced graft copolymer with poor thermal stability. Copyright © 2004 Society of Chemical Industry     

Involvement of amino acid residues 423-429 of human protein S in binding to C4b-binding protein

Human protein S binds to C4b-binding protein (C4BP) both in plasma and in a system using purified proteins. Amino acid residues 420-434 of the first disulfide loop of the sex hormone binding globulinlike domain of protein S are involved in the interaction of protein S with C4BP. To define the involvement of specific polar amino acids within residues 420-434, we studied in parallel synthetic protein S peptides and recombinant protein S variants containing the same amino acid replacements, K423E, E424K, Q427E and K429E. Synthetic peptide analogs of peptide PSP-420 (residues 420-434) were assayed for binding C4BP and as inhibitors of complex formation. The PSP-420 peptide and the analogous peptide with the substitution E424K, but not the peptides containing the substitutions K423E and K429E, were able to bind C4BP. Recombinant proteins with mutations of K423E, Q427E and K429E showed reduced affinity for C4BP compared to plasma protein S, recombinant wild type protein S, or E424K-protein S. These results suggest that Lys-423, Gln-427 and Lys-429 of protein S are important for normal binding to C4BP. The anti-protein S monoclonal antibody LJ-56, raised against peptide PSP-420, recognizes only free protein S and inhibits complex formation with C4BP. Antibody LJ-56 recognized the E424K and Q427E peptides but not the K423E or K429E peptides. Similarly, the E424K and Q427E protein S mutants were recognized by LJ-56, whereas the K423E and K429E protein S mutants were not recognized. This suggests that both in the peptide PSP-420 and in protein S, Lys-423 and Lys-429 significantly contribute to binding to antibody LJ-56. These results demonstrate that protein S residues 423, 427 and 429, but not residue 424, are involved in binding to both the antibody LJ-56 and to C4BP. When peptides PSP 420 and SL-6 (residues 447-460) with carboxyterminal amide or carboxylate moieties were compared to their ability to inhibit C4BP-protein S complexation, PSP-420-amide was the most potent. This finding together with the other results described here supports the hypothesis that the residues 420 and 434 in protein S provides a major binding site for C4BP.     

Plasma TAFI levels influence the clot lysis time in healthy individuals in the presence of an intact intrinsic pathway of coagulation

Thrombin Activatable Fibrinolysis Inhibitor (TAFI) is a recently identified fibrinolysis inhibitor in plasma, that when converted to an enzyme potently attenuates fibrinolysis. It is activated by relatively high concentrations of thrombin that exceed the thrombin concentration required for fibrin formation. These high concentrations of thrombin are generated by the intrinsic pathway via activation of factor XI by thrombin. The down regulation of fibrinolysis by TAFI can be measured in a clot lysis assay. When the clot lysis times of healthy individuals were determined, large inter-individual differences were observed. To determine if differences in concentration of TAFI explain the variation in clot lysis between individuals, specific assays were developed for the measurement of TAFI antigen and activity in plasma. In normal plasma, there was a dose-dependent relationship between TAFI antigen and TAFI activity. There was also a correlation between clot lysis time and plasma TAFI antigen, indicating that the amount of TAFI that is activated during the clot lysis assay, is dependent on the concentration of TAFI. In the plasmas of 20 healthy individuals, clot lysis times, TAFI antigen and TAFI activity were determined. Both TAFI antigen and TAFI activity showed a significant correlation with the clot lysis time. No correlation between TAFI antigen and clot lysis time was found when the clot lysis time was determined in the presence of an antibody blocking the factor XI feedback loop. These results indicate that plasma TAFI levels influence the clot lysis time in healthy individuals in the presence of an intact intrinsic pathway of coagulation.     

Familial neuroblastoma: report of a kindred with later age at diagnosis

The first reported effective adjuvant combination regimen for patients with operable breast cancer comprised oral cyclophosphamide (C) days 1-14 with intravenous methotrexate (M) and fluorouracil (F) on days 1 and 8, repeated every 28 days ('classical' CMF). These drugs have since been extensively used with or without endocrine therapies and/or other cytotoxics, as well as with radiation therapy to the chest wall yielding conflicting results. Although doses and schedules have varied widely, the combination of these three drugs has been generically referred to as CMF. Evidence exists that reducing the dose and/or altering the schedule of CMF ('modified' CMF) have compromised its efficacy in metastatic breast cancer. Reduction below standard dose of a similar regimen also gave inferior results in the adjuvant setting. In fact, the recently reported improved outcome of adding radiation therapy to CMF was only demonstrated in comparisons with a 'modified' CMF. Furthermore, trials in women with estrogen receptor-positive breast cancer, which did not demonstrate any significant benefit for the addition of adjuvant CMF to tamoxifen compared with tamoxifen alone, also used 'modified' CMF. Therefore, adherence to the 'classical' dose and schedule is recommended when CMF is used in adjuvant therapy.