Construction and characterization of a radio-iodinatable mutant of recombinant human CD4

Drug-selected intrachromosomal gene amplification by breakage-fusion-bridge (BFB) cycles is well documented in mammalian cells, but factors governing this mechanism are not clear. Here, we show that only some clastogenic drugs induce drug resistance through intrachromosomal amplification. We strictly correlate triggering of BFB cycles to induction of fragile site expression. We demonstrate a dual role for fragile sites in intrachromosomal amplification: a site telomeric to the selected gene is involved in initiation, while a centromeric site defines the size and organization of early amplified units. The positions of fragile sites relative to boundaries of amplicons found in human cancers support the hypothesis that fragile sites play a key role in the amplification of at least some oncogenes during tumor progression.     

Modulation of excitatory synaptic transmission in locus coeruleus by multiple presynaptic metabotropic glutamate receptors

Metabotropic glutamate receptors have been implicated in modulation of synaptic transmission in many different systems. This study reports the effects of selective activation of metabotropic glutamate receptors on synaptic transmission in intracellularly recorded locus coeruleus neurons in brain slice preparations. Perfusion of either L-2-amino-4-phosphonobutyric acid (L-AP4; 0.1-500 microM) or (+/-)-1-aminocyclopentane-trans-1,3,dicarboxylic acid (t-ACPD; 0.1-500 microM) caused a depression of excitatory postsynaptic potentials in a dose-dependent fashion to about 70% inhibition. Both agonists exerted their effects at relatively low concentrations with estimated EC50s of 2.6 microM and 11.5 microM for L-AP4 and t-ACPD, respectively. This inhibition was not observed with the potent group I metabotropic glutamate receptor agonist (RS)-3,5-dihydroxyphenylglycine (DHPG; 100 microM). Conversely, (R)-4-carboxy-3-hydroxyphenyl-glycine (4C-3H-PG), a group I antagonist/group II agonist, and 2R,4R-4-aminopyrrolidine-2,4-dicarboxylate (APDC), a novel and specific group II agonist, also caused an inhibition of excitatory postsynaptic potentials. Both t-ACPD and L-AP4 produced an increase in paired-pulse facilitation, and failed to change the locus coeruleus response to focally applied glutamate, indicating a presynaptic locus of action. The L-AP4 inhibition was antagonized by (S)-amino-2-methyl-4-phosphonobutanoic acid (MAP4: group III antagonist) but not by (RS)-alpha-methyl-4-carboxyphenylglycine [(RS)-MCPG; mixed antagonist], suggesting that this agonist acts through a type 4 metabotropic glutamate receptor. Conversely, t-ACPD was antagonized by MCPG and by ethyl glutamate (group II antagonist), but not by aminoindan dicarboxylic acid (AIDA; group I antagonist) or MAP4, suggesting that this agonist acts on a type 2 or 3 metabotropic glutamate receptor. Taken together, these results suggest that two pharmacologically distinct presynaptic metabotropic glutamate receptors function in an additive fashion to inhibit excitatory synaptic transmission in locus coeruleus neurons. These receptors may be involved in a feedback mechanism and as such may function as autoreceptors for excitatory amino acids.     

Absorbable pulmonary artery band

Absorbable pulmonary artery banding may be a useful method of avoiding further operation or angioplasty in patients whose underlying lesion has a natural history of resolution. We report 2 cases of absorbable pulmonary artery banding using braided Dexon. In both cases the bands functioned well initially and were completely resorbed after 2 years and 6 months, respectively.     

Experimental evaluation of a new device for percutaneous transrenal ureteral occlusion

PURPOSE: To develop a device for percutaneous transrenal ureteral occlusion. MATERIALS AND METHODS: The device was a double-body Gianturco-R?sch biliary stent constrained at the junction of the two stents to create an hourglass shape. One stent was coated with silicone. One device was percutaneously placed in each of nine pigs through a 9-F Teflon sheath. Urographic and hematologic follow-up was performed for up to 12 weeks. RESULTS: Seven pigs showed immediate, complete ureteral occlusion, and two pigs exhibited persistent incomplete high-grade obstruction. All animals exhibited varying degrees of hydronephrosis and hydroureter. No device migration was noted. Minor complications were encountered during device placement in three pigs. Mucosal folds and villus-like projections that arose from the lamina propria protruded into the lumen of the ureter at the cranial end of the covered stent and around the wire of the caudal stent. Varying degrees of mural inflammation and edema were noted. CONCLUSION: Transrenal ureteral occlusion with the described device appears to be a viable method for treating urinary fistulas.     

Immune complex-dissociated p24 antigen in congenital or perinatal HIV infection: role in the diagnosis and assessment of risk of infection in infants

Both suppressor oncogene and proliferative activity are believed to indicate colon cancer risk. The retinoblastoma (Rb) gene is a suppressor oncogene affecting cell differentiation. Retinoblastoma gene inactivation is associated with tumour development. However, the relation of the Rb protein to cell proliferation and colon tumour formation is unknown. Retinoblastoma protein quantity was correlated with proliferative activity in flat, unaffected mucosa specimens from 36 cancer patients, 21 non-cancer control subjects and in 29 tumour tissue samples from cancer patients. Nuclear Rb protein was measured by using automated CAS-200 image analysis of monoclonal antibody labelled frozen sections from fresh, surgically removed tissue. All colon cells within 15 whole crypts were imaged. Proliferative activity was also measured by using analysis with Ki-67 monoclonal antibody. Retinoblastoma protein content correlated directly with proliferative activity in flat mucosa of non-cancer control subjects (r = 0.63; P < 0.001; n = 21). A significant correlation was also found in flat mucosa specimens of non-metastatic (Duke's stages A and B) cancer patients (r = 0.52; P < 0.01; n = 22). However, Rb protein did not correlate with proliferation in flat mucosa from metastatic (Duke's stages C and D) cancer patients (r = 0.03; NS; n = 14) or in cancer tissue (r = 0.068; NS; n = 29). Mucosal Rb protein in the colon normally increases as proliferation increases. Dissociation between Rb protein and colon proliferation may occur in flat mucosa in patients with a higher risk of metastatic tumour growth. Future studies comparing Rb protein quantity and proliferative activity may help identify high-risk colon cancer patients.     

Variability in cytotoxicity and fluoride release of resin-modified glass-ionomer cements

New-generation glass-ionomer cements contain resin to improve their restorative properties. These resin-modified glass-ionomer cements vary considerably in their chemistry, which could result in corresponding variability in their physical and biological properties. This study investigated the cytotoxicity and the fluoride release of two resin-modified glass ionomers, a conventional glass-ionomer cement, and a resin composite. Samples were prepared and extracted in distilled water for 1, 4, and 7 days; eluates were filtered and tested by means of 3T3 mouse fibroblasts. Cytotoxicity (MTT assay) values were low for all materials and extraction times, indicating minimal cytotoxicity of all materials (less than 30% inhibition). Cytotoxicity of one resin-modified glass ionomer was significantly higher than for the other materials (p < 0.001). One resin-modified glass ionomer and the conventional glass-ionomer cement released significantly more fluoride at each time interval (p < 0.001) than the other resin-modified glass-ionomer cement and the resin composite. Fluoride release and cytotoxicity were correlated (r2 = 0.60; p < 0.001), although the fluoride release does not account for the cytotoxicity observed. Cytotoxicity and fluoride release suggest that one hybrid behaved more like a conventional glass ionomer, and the other like a resin composite. These differences may have implications for material selection in specific clinical situations.     

The impact of ischemic heart disease on main pulmonary artery blood flow patterns: a comparison between magnetic resonance phase velocity mapping and transesophageal color Doppler

OBJECTIVE: To give a detailed evaluation on main pulmonary artery blood velocity patterns, in patients with ischemic heart disease and to provide recommendations for pulsed Doppler sample volume placement, in order to optimize cardiac output estimation. METHODS: Using magnetic resonance phase and esophageal color Doppler velocity mapping in 12 patients with ischemic heart disease and undergoing coronary artery by-pass grafting, very similar data on pulmonary artery blood velocity patterns were provided for comparison with each other. RESULTS: Peak blood velocities were located in the inferior half of the main pulmonary artery cross-sectional area. Early after peak systole the highest velocities shifted towards the superior/left (major curvature) with a simultaneous decrease in velocities inferiorly. The velocity decrease further evolved into retrograde flow to the inferior/right (minor curvature). This feature was significantly enhanced compared to earlier findings in healthy volunteers. The mean temporal blood velocity profiles were asymmetrically skewed, thereby giving unreliable cardiac output estimates based on single point Doppler blood velocity recordings. The error incurred may amount to more than 100% in extreme cases. According to our data, optimal assessment of cardiac output should be based on multiple sample volumes placed along the inferior/right to superior/left diameter. CONCLUSIONS: MR-phase velocity mapping and multiplane transesophageal color Doppler recordings provided similar blood velocity patterns in patients with ischemic heart disease. The skewness of the mean temporal blood velocity profile is enhanced compared with healthy subjects, resulting in error in the assessment of CO by means of pulsed Doppler echocardiography. By using multiple Doppler sample volumes, the error can be minimized.     

School-screening for scoliosis. A prospective epidemiological study in northwestern and central Greece

A two-year prospective study was done to assess the prevalence and distribution of various parameters associated with scoliosis in schoolchildren in northwestern and central Greece. A total of 82,901 children (41,939 boys and 40,962 girls) who were nine to fourteen years old were screened for scoliosis. Five thousand eight hundred and three children had clinical signs of scoliosis and, of these, 4185 were referred for posteroanterior radiographs (to be made with the patient standing) because they had a positive result on the forward-bending test (a difference of more than five millimeters between the two sides of the torso as measured in the thoracic or thoracolumbar region with use of a ruler and a level plane) at the time of a second screening. The prevalence of scoliosis (defined as a curve of 10 degrees or more) was 1.7 per cent (1436 of 82,901 children), and most of the curves (1255; prevalence, 1.5 per cent) were small (10 to 19 degrees). The ratio of boys to girls was 1:2.1 over-all but varied according to the magnitude of the curve (1:1.5 for curves of less than 10 degrees, 1:2.7 for curves of 10 to 19 degrees, 1:7.5 for curves of 20 to 29 degrees, 1:5.5 for curves of 30 to 39 degrees, and 1:1.2 for curves of 40 degrees or more). Thoracolumbar curves were the most common type of curve identified, followed by lumbar curves; specifically, of the 1436 children who had a curve of at least 10 degrees, 493 (34.3 per cent) had a thoracolumbar curve, 475 (33.1 per cent) had a lumbar curve, 261 (18.2 per cent) had a thoracic curve, and 207 (14.4 per cent) had a double curve. Although most (753) of these curves were to the left, the left:right ratio varied according to the location of the apex of the curve (1:3.1 for thoracic curves, 2.0:1 for thoracolumbar curves, and 3.2:1 for lumbar curves). The cost of the screening process was negligible (estimated at thirty cents per child); however, the decreased number of operative procedures performed in children from the geographical area of our University Hospital, the identification of a large number of previously undiagnosed curves (eleven of which were treated operatively and 170 of which were treated with a brace), and the identification of children who were at high risk for progression were considered important benefits of the school-screening program.