Molecular cloning and tissue expression analysis of the beta subunit of elongation factor 1 in the mouse.
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Biomedical subjects
Publications and source records attributed to S Zhao.
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Transforming growth factor-beta (TGF-beta) sequestered in dentine matrix has an important role in dental tissue repair after injury and its exposure at sites of injury may stimulate tertiary dentinogenesis. This study aimed to investigate the expression of TGF-beta isoforms in mature human dentine matrix and the ability of chemical treatments to expose TGF-beta on the cut surface of dentine using gold immunolabelling and subsequent scanning electron microscopy examination. TGF-beta1 was the only isoform that could be detected in human dentine and the nature of the chemical treatment of the tissue influenced its detection. EDTA treatment provided good exposure of TGF-beta1 on the dentine surface, whilst citric acid and sodium hypochlorite treatments revealed lesser amounts of this isoform. Only minimal staining for TGF-beta1 was observed in samples treated with phosphate-buffered saline. TGF-beta2 and -beta3 could not be detected in the specimens with any of the treatments. This study suggests that TGF-beta1 is the only TGF-beta isoform expressed by human odontoblasts to be sequestered in dentine implying that differences in isoform-extracellular matrix interactions may exist. Information on chemical treatment of tissue specimens for immunostaining may provide a useful basis for selection of tissue preparation techniques for clinical restorative treatment procedures to facilitate TGF-beta mediated reparative processes at sites of dental injury.
Latexin, a carboxypeptidase A inhibitor, is expressed in a cell type-specific manner in both central and peripheral nervous systems in the rat. It is used as a molecular marker for the regional specification of the neocortex. In this study, a cDNA was isolated from a human fetal brain cDNA library. The cDNA (LXN) contains an open reading frame encoding 222 amino acids. The comparison between the deduced amino acid sequences of LXN and latexins of rat and mouse revealed high sequence identity (84.2 and 84.7%, respectively). Northern blot analysis showed that LXN was expressed as a transcript of 1.3 kb in 15 out of 16 tissues examined, except in peripheral blood leukocyte. The expression levels were high in heart, prostate, ovary, kidney, pancreas, and colon, moderate or low in other tissues including brain. It is noteworthy that the tissue distribution of human LXN differs greatly to that of its homologue in the model animal, rat latexin. In addition, the LXN gene contains at least 6 exons and spans 5.9 kb according to the genomic sequence of the clone RP11-79M21 and the gap sequence cloned in this paper. LXN was assigned to 3q25-q26.2 according to the position of the marker SHGC-35682 found adjacent to LXN gene.
RalGDS is a guanine nucleotide dissociation stimulator for Ral, which is a member of the Ras GTPase superfamily that regulates cellular proliferation, differentiation and transformation by mediating multiple signal transduction pathways. RalGDS can specifically promote the conversion from an inactive GDP-bound state to an active GTP-bound state for Ral. The cDNA of human RalGDS has been cloned recently. In this paper, by comparison between the gene's genomic and cDNA seqence, we determined the structure of the gene, which showed that the reported human RalGDS transcribed from 18 exons. Furthermore, a novel variant of RalGDS that codes for a protein with a different N-terminus was cloned and identified. Northern hybridization revealed that the novel transcript was of 6.0 kb in length while the transcript previously reported is of 4.0 kb. Both transcripts were ubiquitously expressed in human adult tissues examined, albeit with different amounts. In addition, this novel transcript was proved to be caused by employment of a new exon, designated as exon 1a, instead of the one, designated as exon 1b, in the reported cDNA. Thus, the RalGDS gene consists of at least 19 exons and spanned a 44 kb region. The length between exon 1a and exon 2 was 33 kb, while the length between exon 1b and exon 2 was 8.8 kb.
Cells regulate phospholipase D (PLD) activity in response to numerous extracellular signals. Here, we investigated the involvement of PLD activity in transforming growth factor-beta (TGF-beta1)-mediated growth inhibition of epithelial cells. TGF-beta1 inhibits the growth of MDCK, Mv1Lu, and A-549 cells. In the presence of 0.4% butanol, TGF-beta1 induces an increase in the formation of phosphatidylbutanol, a unique product catalyzed by PLD. TGF-beta1 also induces an increase in phosphatidic acid (PA) level in A-549 and MDCK cells. TGF-beta1 induces an increase in the levels of DAG labeled with [3H]-myristic acid in A-549 and MDCK cells but not in Mv1Lu cells. No increase of DAG was observed in cells prelabeled with [3H]-arachidonic acid. The data presented suggest that PLD activation is involved in the TGF-beta1-induced cell growth inhibition.
The genetic diversities and relationships of four Chinese indigenous pig breeds and one Australian commercial pig breed have been evaluated using 27 microsatellites recommended by the International Society of Animal Genetics (ISAG) and the Food and Agriculture Organization (FAO). The allele frequencies, effective numbers of alleles and the polymorphic information content have been calculated. Nei's standard genetic distances have been used to construct a UPGMA dendrogram, which has been evaluated by the Bootstrap test. The utility of microsatellites for evaluating genetic diversity of pigs is discussed.
An extremely rare case of unilateral gonadoblastoma with mixed germ cell tumor arising in the ovary of a 27-year-old woman with 46,XX karyotype and two successful pregnancies is reported. The mixed germ cell tumor was composed of choriocarcinoma, embryonal carcinoma, yolk sac tumor, immature teratoma and dysgerminoma. The patient has been well, without evidence of disease for over 10 years since her first surgery and adjuvant chemotherapy.
Different types of biodegradable membranes have become available for guided tissue regeneration. The purpose of this study was to evaluate histologically three different biodegradable membranes (Bio-Gide, Resolut and Vicryl) and one non-biodegradable membrane (expanded polytetrafluoroethylene/e-PTFE) implanted subcutaneously in rats. Five subcutaneous pouches were created in each of 24 rats. One of the four test membranes was randomly placed in each of the four pouches and one pouch was left empty to serve as a control. Histological evaluation was performed after 4, 10 and 21 days which demonstrated that e-PTFE was well tolerated and encapsulated by a fibrous connective tissue capsule. There was capsule formation around Resolut and Vicryl and around Bio-Gide in the early phase there was a wide inflammatory zone already. e-PTFE and Vicryl were stable materials while Resolut and Bio-Gide fragmented in the early phase. In the late phase Vicryl was surrounded by an increasing amount of multinucleated macrophages and a thin capsule, whilst around Resolut and Bio-Gide a strong foreign body reaction was observed. Also granuloma formation was noted around the fragmented Resolut material in its capsule and a mild inflammatory reaction surrounding Bio-Gide within its thin capsule.
Three microsatellite loci on porcine chromosome 12 were ordered by single sperm typing to expand the limited genetic map of this region. Individual sperm cells from a Chinese indigenous Qingping boar triply heterozygous at SW874, SW1350 and SW1553 were amplified using PEP and heminesting primer design at each locus. Analysis of the sperm typing data by the SPERM.FOR program showed that the most likely order was SW1553-SW1350-SW874.
BACKGROUND: Before universal infant immunization against hepatitis B virus (HBV) in 1986 China was a region endemic for HBV infection. The prevalence of HBV infection in the population was about 60% and the proportion of chronic HBV carriers around 10%. These HBV carriers could progress to chronic hepatitis B, cirrhosis, and primary hepatocellular carcinoma. Since 1976, large-scale sero-surveys of HBV infection have been carried out and a lot of data have been collected. METHOD: This paper describes a mathematical model developed to predict the dynamics of HBV transmission and to evaluate the long-term effectiveness of the vaccination programme. We used a compartment model expressed by a set of partial differential equations based on the characteristics of HBV infection. RESULTS: All parameters, expressed in the model as a non-linear function of age and time since vaccination, were estimated using sero-survey data. The model fits well with both pre-vaccination and post-vaccination sero-surveys. The observed and estimated age-specific prevalence rates of HBV infection and HBV carriage agree with each other. According to our model, if all newborns are vaccinated according to schedule, the rate of HBV carriage will decline sharply over time to 0.2% in 70 years. By then, the ratio of acute hepatitis B will be less than 0.5% and the ratio of chronic hepatitis B will be around 5%. CONCLUSIONS: The results suggest that HBV infection in China can be controlled in just one generation, and eventually eliminated. Our model shows that vaccination coverage is the most important indicator for the elimination of HBV transmission. The higher the vaccination coverage, the better the long-term effectiveness of immunization. Thus, the key to controlling and eliminating HBV transmission in China is to find ways to immunize all infants throughout the country, especially in poor, rural areas.
X-ray computed tomography (CT) is in transition from fan-beam to cone-beam geometry. For cone-beam volumetric imaging, reduction of radiation exposure remains an important issue. Because the wavelet approach was shown to be effective and flexible for two-dimensional (2-D) local region reconstruction, we are motivated to perform wavelet local CT in cone-beam geometry. In this paper, we formulate the Feldkamp cone-beam reconstruction from the wavelet perspective, derive both full-scan and half-scan Feldkamp-type formulas for either global or local reconstruction, and demonstrate the feasibility and utility in synthetic and real data. It is found that using the wavelet Feldkamp approach, a three-dimensional (3-D) region of interest (ROI) can be reconstructed with neither severe image artifacts nor any significant constant bias in our simulation and experiments.
Traditional computed tomography (CT) reconstructions of total joint prostheses are limited by metal artifacts from corrupted projection data. Published metal artifact reduction methods are based on the assumption that severe attenuation of X-rays by prostheses renders corresponding portions of projection data unavailable, hence the "missing" data are either avoided (in iterative reconstruction) or interpolated (in filtered backprojection with data completion; typically, with filling data "gaps" via linear functions). In this paper, we propose a wavelet-based multiresolution analysis method for metal artifact reduction, in which information is extracted from corrupted projection data. The wavelet method improves image quality by a successive interpolation in the wavelet domain. Theoretical analysis and experimental results demonstrate that the metal artifacts due to both photon starving and beam hardening can be effectively suppressed using our method. As compared to the filtered backprojection after linear interpolation, the wavelet-based reconstruction is significantly more accurate for depiction of anatomical structures, especially in the immediate neighborhood of the prostheses. This superior imaging precision is highly advantageous in geometric modeling for fitting hip prostheses.
Myelotoxicity is the main factor which decides the maximum tolerated dose (MTD) in radioimmunotherapy (RIT). Since bone marrow is mostly irradiated from blood radioactivity, enhancing the clearance of unbound circulating radiolabeled antibody is important to reduce myelotoxicity and to increase the MTD. We applied the avidin chase method, which was devised to obtain high tumor-to-background ratios in tumor-targeting, to RIT of experimental liver micrometastases and evaluated its influence on the side effects and therapeutic outcome. Seven days after intrasplenic injection of human colon cancer LS174T cells, nude mice were intravenously injected with biotinylated (131)I-labeled anti-CEA monoclonal antibody (MAb) (24 - 38 microg, 11.1 MBq). Mice of the chase group then received an intravenous injection of avidin twice (24 and 30 h, 72 - 115 microg each). Biodistribution, side effects (white blood cell counts and body weight change), and short- and long-term therapeutic effects were determined. Avidin chase markedly accelerated the clearance of radiolabeled MAb from the blood (P < 0.0001) and normal tissues, resulting in milder leukocytopenia and body weight loss, both of which recovered earlier than in the non-chase group (P < 0.01). The tumor uptake of radiolabeled MAb was also decreased by avidin chase, but the metastases-to-background ratios were increased. Avidin chase gave the therapeutic gain ratio of 1.89. Treated groups with and without avidin chase showed significant therapeutic effects compared to the non-treated group. There was no significant difference in the therapeutic effects between the two treated groups. Avidin chase effectively reduced the side effects of RIT and should increase the MTD.
The entorhinal cortex gives rise to a massive projection to the hippocampus and fascia dentata. In the rat, this projection forms early in development with first entorhinal axons reaching the hippocampus around embryonic day (E) 17. From the very beginning, the entorhinal axons recognize their appropriate termination zones in the hippocampus proper and fascia dentata, i.e., stratum lacunosum-moleculare and the outer molecular layer of the dentate. This is remarkable, because at the time of entorhinal fiber ingrowth, the definitive target cells of entorhinal axons, pyramidal cells and granule cells, are not yet fully developed, and the majority of their distal dendritic tips have not yet reached these layers. This raises the question as to the cellular and molecular signals guiding the entorhinal axons to and keeping them in their target layers. Here we hypothesize that early generated Cajal-Retzius (CR) cells located in stratum lacunosum-moleculare and the outer molecular layer of the dentate, and in particular their axons projecting to the entorhinal cortex, provide a template that is used by the entorhinal axons to find their target layers in the hippocampus.
Fluoroquinolone-resistant avian Escherichia coli isolates from northern Georgia were investigated for gyrA and parC mutations. All isolates contained a mutation in GyrA replacing Ser83 with Leu; seven isolates also contained mutations replacing Asp87 with either Gly or Tyr. Random amplified polymorphic DNA analysis revealed that quinolone-resistant E. coli isolates were genetically diverse.
The TyrR protein of Escherichia coli (513 amino acid residues) is the chief transcriptional regulator of a group of genes that are essential for aromatic amino acid biosynthesis and transport. The TyrR protein can function either as a repressor or as an activator. The central region of the TyrR protein (residues 207 to 425) is similar to corresponding polypeptide segments of the NtrC protein superfamily. Like the NtrC protein, TyrR has intrinsic ATPase activity. Here, we report that TyrR possesses phosphatase activity. This activity is subject to inhibition by L-tyrosine and its analogues and by ATP and ATP analogues. Zinc ion (2 mM) stimulated the phosphatase activity of the TyrR protein by a factor of 57. The phosphatase-active site of TyrR was localized to a 31-kDa domain (residues 191 to 467) of the protein. However, mutational alteration of distant amino acid residues at both the N terminus and the C terminus of TyrR altered the phosphatase activity. Haemophilus influenzae TyrR (318 amino acid residues), a protein with a high degree of sequence similarity to the C terminus of the E. coli TyrR protein, exhibited a phosphatase activity similar to that of E. coli TyrR.
Florfenicol, a veterinary fluorinated analog of thiamphenicol, is approved for treatment of bovine respiratory pathogens in the United States. However, florfenicol resistance has recently emerged among veterinary Escherichia coli isolates incriminated in bovine diarrhea. The flo gene, which confers resistance to florfenicol and chloramphenicol, has previously been identified in Photobacterium piscicida and Salmonella enterica serovar Typhimurium DT104. The flo gene product is closely related to the CmlA protein identified in Pseudomonas aeruginosa. The cmlA gene confers nonenzymatic chloramphenicol resistance via an efflux mechanism. Forty-eight E. coli isolates recovered from calves with diarrhea, including 41 that were both chloramphenicol and florfenicol resistant, were assayed for the presence of both flo and cmlA genes. Forty-two of the 44 isolates for which florfenicol MICs were > or =16 microg/ml were positive via PCR for the flo gene. All E. coli isolates for which florfenicol MICs were < or =8 microg/ml were negative for the flo gene (n = 4) Twelve E. coli isolates were positive for cmlA, and chloramphenicol MICs for all 12 were > or =32 microg/ml. Additionally, eight isolates were positive for both flo and cmlA, and both florfenicol and chloramphenicol MICs for these isolates were > or =64 microg/ml. DNA sequence analysis of the E. coli flo gene demonstrated 98% identity to the published GenBank sequences of both serovar Typhimurium flo(St) and P. piscicida pp-flo. The flo gene was identified on high-molecular-weight plasmids of approximately 225 kb among the majority of florfenicol-resistant E. coli isolates. However, not all of the florfenicol-resistant E. coli isolates tested contained the large flo-positive plasmids. This suggests that several of the E. coli isolates may possess a chromosomal flo gene. The E. coli flo gene specifies nonenzymatic cross-resistance to both florfenicol and chloramphenicol, and its presence among bovine E. coli isolates of diverse genetic backgrounds indicates a distribution much wider than previously thought.
The relationship between changes in the muscle total adenine nucleotide pool (TAN = ATP + ADP + AMP) and IMP during and after 30 s of sprint cycling was examined. Skeletal muscle samples were obtained from the vastus lateralis muscle of seven untrained men (23. 9 +/- 2.3 yr, 74.4 +/- 3.6 kg, and 55.0 +/- 2.9 ml. kg(-1). min(-1) peak oxygen consumption) before and immediately after exercise and after 5 and 10 min of passive recovery. The exercise-induced increase in muscle IMP was linearly related to the decrease in muscle TAN (r = -0.97, P < 0.01), and the slope of this relationship (-0.83) was not different from 1.0 (P > 0.05), indicating a 1:1 stoichiometric relationship. This interpretation must be treated cautiously, because all subjects displayed a greater decrease in TAN compared with the increase in IMP content, and the TAN + IMP + inosine + hypoxanthine content was lower (P < 0.05) immediately after exercise compared with during rest. During the first 5 min of recovery, the increase in TAN was not correlated with the decrease in IMP (r = -0.18, P > 0.05). In all subjects, the magnitude of TAN increase was higher than the magnitude of IMP decrease over this recovery period. In contrast, the increase in TAN was correlated with the decrease in IMP throughout the second 5 min of recovery (r = -0.80, P < 0.05), and it was a 1:1 stoichiometric relationship (slope = -1.12). These data indicate that a small proportion of the TAN pool was temporarily lost from the muscle purine stores during sprinting but was rapidly recovered after exercise.