Searched for: school:SOM
Department/Unit:Cell Biology
Correlation of Web usage and exam performance in a human anatomy and development course
Rizzolo, Lawrence J; Aden, Marcus; Stewart, William B
Course materials for a Human Anatomy and Development Course were placed on the World Wide Web (WWW). The materials included a lab manual, lecture notes and slides, faculty-generated atlases, Web links, and examinations. The lab manual, lecture notes, and atlases were also provided as black-and-white hardcopy. The Office of Education assigned students a code name that allowed them to use the Web site and take exams anonymously. Student Web use was tracked and correlated with their performance on the final examination. Overall use patterns revealed that most students used the Web site to prepare for examinations, but not for daily studying. Old examinations were the most accessed documents; lecture notes were the least accessed. The access patterns of the students with top 20, middle 20 (closest to the mean), and bottom 20 scores on the final examination were compared. In general, there was little difference between the middle and top groups. Students in the bottom group used computer resources significantly less than the other groups. In a second analysis, the 10 students who used the Web site most frequently scored below the mean. The study suggests that interactive exercises will be heavily used, but that the preparation of all course materials for the WWW may not be an efficient use of institutional resources.
PMID: 12203379
ISSN: 0897-3806
CID: 382832
Role of a highly conserved NH(2)-terminal domain of the human parainfluenza virus type 3 RNA polymerase
Malur, Achut G; Choudhary, Suresh K; De, Bishnu P; Banerjee, Amiya K
The RNA polymerase complex of human parainfluenza virus type 3 (HPIV 3), a member of the family Paramyxoviridae, is composed of two virally encoded polypeptides: a multifunctional large protein (L, 255 kDa) and a phosphoprotein (P, 90 kDa). From extensive deduced amino acid sequence analyses of the cDNA clones of a number of L proteins of nonsegmented negative-strand RNA viruses, a cluster of high-homology sequence segments have been identified within the body of the L proteins. Here, we have focused on the NH(2)-terminal domain of HPIV 3 L protein that is also highly conserved. Following mutational analyses within this domain, we examined the ability of the mutant L proteins to (i) transcribe an HPIV 3 minireplicon, (ii) transcribe the viral RNA in vitro using the HPIV 3 nucleocapsid RNA template, and (iii) interact with HPIV 3 P protein. Our results demonstrate that the first 15 amino acids of the NH(2)-terminal domain spanning a highly conserved motif is directly involved in transcription of the genome RNA and in forming a functional complex with the P protein. Substitution of eight nonconserved amino acids within this domain by the corresponding Sendai virus L protein residues yielded mutants with variable transcriptional activities. However, one mutant in which all eight amino acids were replaced with the corresponding residues of Sendai virus L protein failed to both transcribe the minireplicon and interact with HPIV 3 P and the Sendai virus P protein. The possible functional significance of the NH(2)-terminal domain of paramyxovirus L protein is discussed.
PMCID:155155
PMID: 12134015
ISSN: 0022-538x
CID: 1444562
Two-color GFP imaging demonstrates cell-autonomy of GAL4-driven RNA interference in Drosophila
Van Roessel, Peter; Hayward, Neil M; Barros, Claudia S; Brand, Andrea H
PMID: 12324976
ISSN: 1526-954x
CID: 5192732
Matrix-matrix interaction of cartilage oligomeric matrix protein and fibronectin
Di Cesare, Paul E; Chen, Frank S; Moergelin, Matthias; Carlson, Cathy S; Leslie, Michael P; Perris, Roberto; Fang, Carrie
Recent work indicates that cartilage oligomeric matrix protein (COMP) plays an important role in extracellular matrix assembly and matrix-matrix protein interactions. In order to identify the proteins in extracellular matrix that interact with COMP, we used an ELISA-based solid-phase binding assay, which revealed a specific, high-affinity interaction between COMP and fibronectin. This interaction is concentration-dependent and saturable, and appears to occur under physiologically relevant conditions. Electron microscopy after negative staining and fragment binding analysis using the solid-phase assay revealed a predominant binding site for the COMP C-terminal globular domain to a molecular domain approximately 14 nm from the N-terminal domain of fibronectin, which can be inhibited by the presence of a polyclonal antibody specific for the C-terminal heptadecapeptide of COMP. This interaction is further demonstrated in vivo by colocalization of both COMP and fibronectin in the chondrocyte pericellular matrix by laser confocal microscopy of chondrocytes grown in agarose culture, and by appositional and colocalization of these proteins in the growth plate of primates by immunohistochemistry
PMID: 12225811
ISSN: 0945-053x
CID: 72049
Lowering of dietary advanced glycation endproducts (AGE) reduces neointimal formation after arterial injury in genetically hypercholesterolemic mice
Lin, Reigh-Yi; Reis, Ernane D; Dore, Anthony T; Lu, Min; Ghodsi, Newsha; Fallon, John T; Fisher, Edward A; Vlassara, Helen
Restenosis remains a major cause of morbidity and mortality after coronary angioplasty. Injury-induced inflammation, thrombosis, smooth muscle cell (SMC) proliferation, and neointimal formation contribute to restenosis. These events are linked to circulating glucose-derived advanced gycation endproducts (AGE), known to promote cell proliferation, lipid glycoxidation and oxidant stress. This study evaluates the association between dietary AGE content and neointimal formation after arterial injury in genetically hypercholesterolemic mice. Male, 12-week-old, apolipoprotein E-deficient (apoE(-/-)) mice were randomly assigned to receive either a high AGE diet (HAD; AGE=15000 U/mg), or a similar diet with ten-fold lower AGE (LAD; AGE=1500 U/mg). These mice underwent femoral artery injury 1 week later, and were maintained on their diets for an additional 4 weeks. At 4 weeks after injury, significant decrease in neointimal formation was noted in LAD-fed mice. Neointimal area, intima/media ratio, and stenotic luminal area (LA) were less pronounced in the LAD group than the HAD group (P<0.05). These quantitative differences were associated with a marked reduction ( approximately 56%) of macrophages in the neointimal lesions, as well as an obvious reduction of SMC content of LAD-fed mice. The reduction of neointimal formation in the LAD mice correlated with a approximately 40% decrease in circulating AGE levels (P<0.0005). Immunohistochemistry also showed a reduced ( approximately 1.5-fold) deposition of AGE in the endothelia, SMC, and macrophages in neointimal lesions of LAD-fed mice. These results represent the first evidence in vivo for a causal relationship between dietary AGE and the vessel wall response to acute injury, suggesting a significant potential for dietary AGE restriction in the prevention of restenosis after angioplasty
PMID: 12052477
ISSN: 0021-9150
CID: 37283
Role of the 5' untranslated region (UTR) in the tissue-specific regulation of rat tryptophan hydroxylase gene expression by stress
Chamas, Firas; Sabban, Esther L
There are tissue specific discrepancies in expression of tryptophan hydroxylase (TPH) between the pineal gland and brainstem. TPH mRNA levels in the pineal are much higher than in the brainstem, however, the two tissues contain comparable protein levels. This discrepancy could result from different translation efficiency of two of the TPH mRNA isoforms. Using PCR-based methods, we analyzed the relative expression, in pineal and brainstem, of two TPH mRNA isoforms differing in the length of their untranslated region (5'UTR). The levels of the TPHalpha were found to be 960-fold more abundant than the 51-nucleotide longer TPHbeta, in the pineal. TPHbeta was also detected for the first time in the brainstem, where TPHbeta/TPHalpha was about five-fold higher than in the pineal. To study the role of the different 5'UTRs, each was cloned in-frame upstream of luciferase, and transfected into PC12 cells. Both 5'UTRs enhanced luciferase activity, with TPHbeta 5'UTR being more effective than TPHalpha 5'UTR, indicating selective regulation of translation efficiency. We also examined whether physiological manipulations alter the distribution of the TPH mRNA isoforms. Repeated stress had no effect in pineal, but led to a marked preferential induction of TPHbeta in brainstem. Modulation of TPH gene expression in serotonergic neurons could result from selective and tissue specific regulation of its mRNA isoforms.
PMID: 12153488
ISSN: 0022-3042
CID: 606992
Analysis of twenty-four Gal4 lines in Drosophila melanogaster
Hrdlicka, Lori; Gibson, Matthew; Kiger, Amy; Micchelli, Craig; Schober, Markus; Schock, Frieder; Perrimon, Norbert
PMID: 12324947
ISSN: 1526-954x
CID: 134524
Efficacy of orally delivered cochleates containing amphotericin B in a murine model of aspergillosis
Delmas, G; Park, S; Chen, Z W; Tan, F; Kashiwazaki, R; Zarif, L; Perlin, D S
Cochleates containing amphotericin B (CAMB) were administered orally at doses ranging from 0 to 40 mg/kg of body weight/day for 14 days in a murine model of systemic aspergillosis. The administration of oral doses of CAMB (20 and 40 mg/kg/day) resulted in a survival rate of 70% and a reduction in colony counts of more than 2 logs in lungs, livers, and kidneys. Orally administered CAMB shows promise for the treatment of aspergillosis.
PMCID:127382
PMID: 12121962
ISSN: 0066-4804
CID: 310422
In Vivo Depletion of CD11c(+) Dendritic Cells Abrogates Priming of CD8(+) T Cells by Exogenous Cell-Associated Antigens
Jung, Steffen; Unutmaz, Derya; Wong, Phillip; Sano, Gen-Ichiro; De los Santos, Kenia; Sparwasser, Tim; Wu, Shengji; Vuthoori, Sri; Ko, Kyung; Zavala, Fidel; Pamer, Eric G; Littman, Dan R; Lang, Richard A
Cytotoxic T lymphocytes (CTL) respond to antigenic peptides presented on MHC class I molecules. On most cells, these peptides are exclusively of endogenous, cytosolic origin. Bone marrow-derived antigen-presenting cells, however, harbor a unique pathway for MHC I presentation of exogenous antigens. This mechanism permits cross-presentation of pathogen-infected cells and the priming of CTL responses against intracellular microbial infections. Here, we report a novel diphtheria toxin-based system that allows the inducible, short-term ablation of dendritic cells (DC) in vivo. We show that in vivo DC are required to cross-prime CTL precursors. Our results thus define a unique in vivo role of DC, i.e., the sensitization of the immune system for cell-associated antigens. DC-depleted mice fail to mount CTL responses to infection with the intracellular bacterium Listeria monocytogenes and the rodent malaria parasite Plasmodium yoelii
PMCID:3689299
PMID: 12196292
ISSN: 1074-7613
CID: 32272
Bmp signaling is required for development of primary lens fiber cells
Faber, Sonya C; Robinson, Michael L; Makarenkova, Helen P; Lang, Richard A
We have investigated the role of Bmp signaling in development of the mouse lens using three experimental strategies. First, we have shown that the Bmp ligand inhibitor noggin can suppress the differentiation of primary lens fiber cells in explant culture. Second, we have expressed a dominant-negative form of the type 1 Bmp family receptor Alk6 (Bmpr1b -- Mouse Genome Informatics) in the lens in transgenic mice and shown that an inhibition of primary fiber cell differentiation can be detected at E13.5. Interestingly, the observed inhibition of primary fiber cell development was asymmetrical and appeared only on the nasal side of the lens in the ventral half. Expression of the inhibitory form of Alk6 was driven either by the alpha A-cystallin promoter or the ectoderm enhancer from the Pax6 gene in two different transgenes. These expression units drive transgene expression in distinct patterns that overlap in the equatorial cells of the lens vesicle at E12.5. Despite the distinctions between the transgenes, they caused primary fiber cell differentiation defects that were essentially identical, which implied that the equatorial lens vesicle cells were responding to Bmp signals in permitting primary fiber cells to develop. Importantly, E12.5 equatorial lens vesicle cells showed cell-surface immunoreactivity for bone-morphogenetic protein receptor type 2 and nuclear immunoreactivity for the active, phosphorylated form of the Bmp responsive Smads. This indicated that these cells had the machinery for Bmp signaling and were responding to Bmp signals. We conclude that Bmp signaling is required for primary lens fiber cell differentiation and, given the asymmetry of the differentiation inhibition, that distinct differentiation stimuli may be active in different quadrants of the eye.
PMID: 12117821
ISSN: 0950-1991
CID: 2329972