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R Luo

Publications and source records attributed to R Luo.

At least 37 records · Page 2Linked to original sources

Avian transitin expression mirrors glial cell fate restrictions during neural crest development.

During development, trunk neural crest cells give rise to three primary classes of derivatives: glial cells, melanocytes, and neurons. As part of an effort to learn how neural crest diversification is regulated, we have produced monoclonal antibodies (MAbs) that recognize antigens expressed by neural crest cells early in development. One of these, MAb 7B3 (7B3), was found to recognize an avian transitin-like protein by co-immunostaining with a series of transitin-specific monoclonal antibodies and by Western blot analysis. In neural crest cell cultures, we found that 7B3 initially recognizes the majority of neural crest cells as they emerge from the neural tube. Subsequently, 7B3-immunoreactivity (IR) is progressively restricted to a smaller subpopulation of cells. In fully differentiated trunk neural crest cell cultures, 7B3-IR is expressed only by cells that do not express neuronal markers and lack melanin granules. During development in vivo, 7B3-IR is evident in neural crest cells on the medial, but not the lateral migration pathway, suggesting that it is not expressed by melanocyte precursors. Later, the antigen is detected in non-neuronal, presumptive glial cells in dorsal root ganglia (DRG) and sympathetic ganglia, as well as along ventral roots. Cultures of E5 DRG confirm that 7B3-IR is restricted to non-neuronal cells of ganglia, many of which closely associate with neuronal processes. Therefore, of the three major classes of differentiated trunk neural crest derivatives, 7B3 exclusively recognizes glial cells, including both satellite glia and Schwann cells. Since the pattern of 7B3 expression in vitro mirrors the pattern of glial cell fate-restrictions in the trunk neural crest lineage, and is expressed by neural crest-derived glia in vivo, we conclude that 7B3 is an early pan-glial marker for neural crest-derived glial cells and their precursors.

Animals↗

Genes dependent on zebrafish cyclops function identified by AFLP differential gene expression screen.

Zebrafish cyclops (cyc) encodes a Transforming Growth Factor beta (TGFbeta) signaling factor closely related to mouse Nodal. By comparing amplified fragment length polymorphisms (AFLP) from cyc mutant and wild-type cDNA pools, we devised a differential gene expression screen to isolate genes whose expression is dependent on Cyc signaling. We report two genes not previously described in the zebrafish that were identified using this approach. The first gene, crestin, is expressed predominantly in premigratory and migrating neural crest cells during somitogenesis stages. crestin expression is reduced in cyc mutants initially but recovers by late somitogenesis. The second gene encodes the zebrafish homologue of the calcium-binding protein, calreticulin. Zebrafish calreticulin is highly expressed in the hatching gland and in the floor plate, tissues that are affected in cyc mutants. During gastrulation, calreticulin transcripts are found in the dorsal mesendoderm, in the same cells that express the cyc gene. Expression is reduced in cyc mutants and is abolished by the one-eyed pinhead (oep) mutation that is presumed to prevent Nodal signaling. The identification of calreticulin suggests that a differential screen between wild-type and mutant cDNA is a useful approach to reveal regulation of unexpected gene expression in response to cellular signals. genesis 26:86-97, 2000.

Animals↗

[Optical coherence tomography applied for measurement of nerve fiber layer thickness in normal eyes].

OBJECTIVE: To evaluate the efficiency of optical coherence tomography (OCT) on quantifying retinal nerve fiber layer (RNFL) thickness and its reproducibility and investigate the difference of RNFL thickness between different groups. METHODS: According to their age, 152 eyes of 152 normal subjects were classified into six groups (such as 10 year, 20 year group, etc.), and investigated by OCT to measure the RNFL thickness. Each subject had circular scans around the optic nerve with a circle size of 3.46 mm. The RNFL thickness in different groups was analyzed. Intraclass correlation coefficient (ICC) in fifty eyes of 37 subjects was calculated. RESULTS: The mean RNFL thickness of 152 eyes was (90.09 +/- 10.81) microm, (140.27 +/- 10.60) microm, (85.03 +/- 14.01) microm, (140.27 +/- 9.70) microm and (114.11 +/- 6.08) microm (temporal, superior, nasal, inferior and the mean of the whole layer, respectively). Least significant difference (LSD)-t test was applied for multiple thickness comparisons of the mean and different quadrants among different groups. The results showed there were significant differences between the 60 year group and others except the 50 year group in temporal, superior, inferior quadrants and mean RNFL thickness (P = 0.000 - 0.019), and between each of 10 to 40 year groups and each of 50 to 60 year groups in the mean RFNL thickness (P = 0.000 - 0.026). ICCs of RNFL thickness measured by OCT were 0.78, 0.66, 0.64, 0.52 and 0.74 (temporal, superior, nasal, inferior and mean, respectively). CONCLUSIONS: Normal RNFL thickness can be measured accurately by OCT. There are some differences of RNFL thickness between different groups in normal persons.

Adolescent↗

[Cloning of a new catechol 1,2-dioxygenase gene (tfd C) from Plesiomonas and its expression in the E. coli].

A new catechol-1,2-dioxygenase gene (tfd C) was cloned from the Plesiomonas using the PCR method. Primers were designed according to the reported sequence of Catechol-1,2-dioxygenase (C120) gene from Alcaligenes eutroplus. The amplified fragment contained a 765 bp open reading frame (ORF), encoding a protein of 255 amino acids. The new tfd C gene shared a high homology with the one cloned from Alcaligenes eutroplus, showing only one base difference at 693 site (C-->A) and consequently one amino acid difference at 228 site (P-->T). The ORF was cloned to the plasmid pBluescriptII KS, which was transferred to E. coli JM109 and a positive clone, pBt2G, was then selected. A significant activity of C120 was detected in the positive clone. When the ORF was cloned to the plasmid pET-30a, which was transferred to E. coli BL21(DE3) plysS, the expected 33 kD protein was detected from a positive clone, pET30A, by SDS-PAGE. The C120 is a key enzyme in degrading aromatic pollutants in the environment. In order to use plants to degrade aromatic pollutants, the gene will be introduced into the turfgrass. To express the gene properly in plants, its translation initiation codon was modified from GTG to ATG. A similar activity of C120 was obtained following the modification.

Amino Acid Sequence↗

Chromosomal aberrations in PARP(-/-) mice: genome stabilization in immortalized cells by reintroduction of poly(ADP-ribose) polymerase cDNA.

Depletion of poly(ADP-ribose) polymerase (PARP) increases the frequency of recombination, gene amplification, sister chromatid exchanges, and micronuclei formation in cells exposed to genotoxic agents, implicating PARP in the maintenance of genomic stability. Flow cytometric analysis now has revealed an unstable tetraploid population in immortalized fibroblasts derived from PARP(-/-) mice. Comparative genomic hybridization detected partial chromosomal gains in 4C5-ter, 5F-ter, and 14A1-C1 in PARP(-/-)mice and immortalized PARP(-/-)fibroblasts. Neither the chromosomal gains nor the tetraploid population were apparent in PARP(-/-) cells stably transfected with PARP cDNA [PARP(-/-)(+PARP)], indicating negative selection of cells with these genetic aberrations after reintroduction of PARP cDNA. Although the tumor suppressor p53 was not detectable in PARP(-/-) cells, p53 expression was partially restored in PARP(-/-) (+PARP) cells. Loss of 14D3-ter that encompasses the tumor suppressor gene Rb-1 in PARP(-/-) mice was associated with a reduction in retinoblastoma(Rb) expression; increased expression of the oncogene Jun was correlated with a gain in 4C5-ter that harbors this oncogene. These results further implicate PARP in the maintenance of genomic stability and suggest that altered expression of p53, Rb, and Jun, as well as undoubtedly many other proteins may be a result of genomic instability associated with PARP deficiency.

Animals↗

Poly(ADP-ribose) polymerase upregulates E2F-1 promoter activity and DNA pol alpha expression during early S phase.

E2F-1, a transcription factor implicated in the activation of genes required for S phase such as DNA pol alpha, is regulated by interactions with Rb and by cell-cycle dependent alterations in E2F-1 abundance. We have shown that depletion of poly(ADP-ribose) polymerase (PARP) by antisense RNA expression downregulates pol alpha and E2F-1 expression during early S phase. To examine the role of PARP in the regulation of pol alpha and E2F-1 gene expression, we utilized immortalized mouse fibroblasts derived from wild-type and PARP knockout (PARP-/-) mice as well as PARP-/- cells stably transfected with PARP cDNA [PARP-/-(+PARP)]. After release from serum deprivation, wild-type and PARP-/-(+PARP) cells, but not PARP-/- cells, exhibited a peak of cells in S phase by 16 h and had progressed through the cell cycle by 22 h. Whereas [3H]thymidine incorporation remained negligible in PARP-/- cells, in vivo DNA replication maximized after 18 h in wild-type and PARP-/-(+PARP) cells. To investigate the effect of PARP on E2F-1 promoter activity, a construct containing the E2F-1 gene promoter fused to a luciferase reporter gene was transiently transfected into these cells. E2F-1 promoter activity in control and PARP-/-(+PARP) cells increased eightfold after 9 h, but not in PARP-/- cells. PARP-/- cells did not show the marked induction of E2F-1 expression during early S phase apparent in control and PARP-/-(+PARP) cells. RT - PCR analysis and pol alpha activity assays revealed the presence of pol alpha transcripts and a sixfold increase in activity in both wild-type and PARP-/-(+PARP) cells after 20 h, but not in PARP-/- cells. These results suggest that PARP plays a role in the induction of E2F-1 promoter activity, which then positively regulates both E2F-1 and pol alpha expression, when quiescent cells reenter the cell cycle upon recovery from aphidicolin exposure or removal of serum.

Animals↗

Application of short-term bioassay guided chemical analysis for water quality of agricultural land run-off.

The effect of agricultural land run-off on the water quality of Lake Kojima, Japan, was investigated using a short-term bioassay-guided chemical analysis. Water samples were collected for 1 year starting from June 1995 to June 1996. Toxicity of the dissolved and adsorbed extracts in the water samples was evaluated using the Daphnia immobilization test and the concentrations of pesticides and putative toxic substance in the extracts were determined by high performance liquid chromatography. Most of the dissolved extracts caused immobilization of the test Daphnia magna at low concentrations during the period of paddy pesticide application. Some extracts were found to contain pesticides such as dymron, mefenacet and flutolanil, but their concentrations were too low to have a toxic effect on the daphnia. An unknown toxic compound, Peak C, was isolated from some river water samples, but it produced only a relatively weak toxicity to Daphnia. To better understand the impact of agricultural run-off on a receiving water body, the relationship between the observed toxicity and the concentrations of pesticides and Peak C in the water samples was studied both temporally and spatially.

Agriculture↗

Poly(ADP-ribosyl)ation of p53 during apoptosis in human osteosarcoma cells.

Spontaneous apoptosis in human osteosarcoma cells was observed to be associated with a marked increase in the intracellular abundance of p53. Immunoprecipitation and immunoblot analysis revealed that, together with a variety of other nuclear proteins, p53 undergoes extensive poly(ADP-ribosyl)ation early during the apoptotic program in these cells. Subsequent degradation of poly(ADP-ribose) (PAR), attached to p53 presumably by PAR glycohydrolase, the only reported enzyme to degrade PAR, was apparent concomitant with the onset of proteolytic processing and activation of caspase-3, caspase-3-mediated cleavage of poly(ADP-ribose) polymerase (PARP), and internucleosomal DNA fragmentation during the later stages of cell death. The decrease in PAR covalently bound to p53 also coincided with the marked induction of expression of the p53-responsive genes bax and Fas. These results suggest that poly(ADP-ribosyl)ation may play a role in the regulation of p53 function and implies a regulatory role for PARP and/or PAR early in apoptosis.

Apoptosis↗

Nucleic acid base-pairing and N-methylacetamide self-association in chloroform: affinity and conformation.

A recently developed computational method, 'mining minima', is used to examine the hydrogen-bonding interactions of nucleic acid base-pairs and of the N-methylacetamide homodimer in chloroform. The mining minima algorithm aggressively samples molecular conformations, identifies the most important local minima, and computes their contributions to the overall free energy of the system. Here, the CHARMM 98 parameter set is used for the potential energy and the generalized Born/surface area solvent model is used to account for the influence of the solvent. Good agreement with experiment is obtained for the non-covalent binding affinities of a series of complexes. The computational approach used here is applicable to a range of molecular systems.

Acetamides↗

[Cervical lymph node metastasis and blood metastasis of thyroid cancer].

OBJECTIVE: To evaluate the relevant factors for lymph cervical node metastases and blood metastases of thyroid cancer and to raise the level of diagnosis and treatment. METHODS: 225 cases of thyroid cancer were treated from Jan. 1984 to Dec. 1992. Their clinical characteristics, treatment, and pathologic features were described. RESULTS: The metastasis of thyroid cancer was related to its pathologic and invasive features. Papillary and squamous cell carcinomas were characterized by local lymph metastasis. Their metastasis rate was 44.7%, and 3/3 respectively. Follicular carcinoma showed good differentiation but vessel metastasis with a metastatic rate of 18.2%. The metastasis rate was high and the prognosis poor, in undifferentiated cancer. CONCLUSIONS: Better treatment results may be achieved if proper management of thyroid cancer is based on its pathologic type and extent of local infiltration.

Adenocarcinoma, Follicular↗

Corrective change of retinal thickness measured by optical coherence tomography and histologic studies.

PURPOSE: To evaluate the correlation of retinal thickness between optical coherence tomography (OCT) images and histologic slides. METHODS: Retinal thickness was measured in 16 rabbit retinal histologic slides. The same eyes had been previously measured by OCT for the comparison of results between two methods. Retinal thickness of each OCT image section was measured using both the manually assisted (requiring localization of reflectivity peaks by observer) and automated modes of the computer software. RESULTS: Retinal thickness measured by OCT demonstrated a high degree of correlation with retinal histologic study. The automated method (Cc = 0.66, P < 0.01) was less reliable than the manually assisted one (Cc = 0.84, P < 0.001). The former had an error in 95% confidence interval, ranged between -0.71 and 11.09 microns. The latter had a less error, ranged from -2.99 to 5.13 microns. CONCLUSION: OCT can quantitatively measure the retinal thickness. However, automatical identification of the reflective boundaries by computer may result in errors in some cases. To measure the retinal thickness by manually assisted mode can increase the accuracy.

Animals↗

[A case-control study on diet and gastric cancer].

OBJECTIVE: To study the relationship between consumption of fish sauce, diet, living-habit and risk of gastric cancer. METHODS: A population-based 1:2 matched case-control study was conducted in Changle and Fuqing Cities of Fujian Province during May 1994 to July 1995. Totally, 272 matched pairs were investigated. RESULTS: Risk state single-factor analysis indicated that there were significant associations between occurrence of gastric cancer and over consumption of fish sauce (OR = 2.57), pickled vegetables (OR = 1.41) and sea food (OR = 1.57), low consumption of fresh vegetables (OR = 1.95), citrus fruits (OR = 1.41), other fruits (OR = 1.31) and green tea (OR = 1.72), exposure to moldy cereals (OR = 2.32), taking meals at irregular times (OR = 5.47) and familial history of malignancy (OR = 3.27). No associations between risk of gastric cancer and smoking, alcohol drinking, amount of salft consumption, use of refrigerator were found. Analysis of risk state conditional logistic regression showed that fish sauce, pickled sea food, irregularly taking meals and familial history of malignancy were included in a set of risk factors for gastric cancer, with a summary attributable risk percentage of 75.49%. CONCLUSION: Over consumption of fish sauce, pickled food, moldy cereals, irregularly taking meals and familial history of malignancy may be the local risk factors for high occurrence of gastric cancer, and fresh vegetables and fruits, green tea may have protective effects on it.

Adult↗

[A quantitative measurement of retinal nerve fiber layer thickness by optical coherence tomography in normal Chinese people].

PURPOSE: To measure retinal nerve fiber layer(RNFL) thickness in normal Chinese people for the use of early diagnosis in glaucomatous patients. METHODS: Optical Coherence Tomography(OCT) was used in 111 eyes of 77 Subjects to create cylindrical cross sections of the RNFL around the optic nerve head with scan diameter of 3.46 mm. The differences in RNFL thickness were analyzed with age, sex, either eye. RESULTS: We found a decrease in RNFL thickness with aging especially in the superior quadrant(P < 0.05), but there weren't significant difference between right and left eyes, neither the sex(P > 0.05). The Mean RNFL thickness of different ages(mean +/- s) was as following: the superior quadrant 128.00 +/- 10.31-149.00 +/- 13.17, the inferior quadrant 132.95 +/- 9.54-142.33 +/- 10.60, the overall mean RNFL thickness 109.00 +/- 4.72-114.28 +/- 7.98 microns. CONCLUSION: The preliminary study showed that neither sex nor either eye were associated with significant differences in RNFL thickness. It is necessary to define the normative value by age group on a larger scale.

Adolescent↗

[Optical coherence tomography and its application in ophthalmology].

Optical coherence tomography (OCT) is a new imaging modality that provides cross-sectional tomographs of ocular structure in vivo. OCT system utilizes propagation of light through, and the time-of-flight delay of light reflected back from biological tissue to produce false-color images of microstructure. OCT has superior spatial resolution (10 microns) than other optical imaging instruments, and maybe achieved at arbitrary penetration depth in transparent tissue with a noncontact, noninvasive measurement. Such features make OCT useful for the early diagnosis, differential diagnosis and more sensitive monitoring of a variety of ocular abnormalities including glaucoma, retinal and anterior eye diseases.

Anterior Eye Segment↗

Confronting the problem of interconnected structural changes in the comparative modeling of proteins.

Comparative models of three proteins have been built using a variety of computational methods, heavily supplemented by visual inspection. We consider the accuracy obtained to be worse than expected. A careful analysis of the models shows that a major reason for the poor results is the interconnectedness of the structural differences between the target proteins and the template structures they were modeled from. Side chain conformations are often determined by details of the structure remote in the sequence, and can be influenced by relatively small main chain changes. Almost all of the regions of substantial main chain conformational change interact with at least one other such region, so that they often cannot be modeled independently. Visual inspection is sometimes effective in correcting errors in sequence alignment and in spotting when an alternative template structure is more appropriate. We expect some improvements in the near future through the development of structure-based sequence alignment tools, side chain interconnectedness rotamer choice algorithms, and a better understanding of the context sensitivity of conformational features.

Amino Acid Sequence↗

The role of excitatory amino acids in the generation of spontaneous hippocampal oscillations.

We examined the role of N-methyl-D-aspartate (NMDA) and non-NMDA glutamate receptors in generating spontaneous CA3 field potential oscillations in the hippocampal slice. Non-NMDA EPSPs are responsible for a portion of the spontaneous activity recorded in standard perfusion medium. NMDA receptors are not activated when inhibition is intact, however, recurrent excitation via NMDA receptors accounts for an increasing proportion of the low frequency (2-4 Hz) rhythms produced as inhibition is progressively blocked by low concentrations of the GABAA antagonist, penicillin. Production of these rhythms involves complex interactions among NMDA, non-NMDA and GABAA receptors. NMDA EPSPs can drive the rhythm in the absence of non-NMDA receptors only when inhibition is suppressed by more than 50%. Otherwise non-NMDA EPSPs appear to be necessary to depolarize neurons before NMDA EPSPs can be activated.

2-Amino-5-phosphonovalerate↗