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PubMed2000

Selective inhibition of splicing at the avian sarcoma virus src 3' splice site by direct-repeat posttranscriptional cis elements.

The direct-repeat elements (dr1) of avian sarcoma virus (ASV) and leukosis virus have the properties of constitutive transport elements (CTEs), which facilitate cytoplasmic accumulation of unspliced RNA. It is thought that these elements represent binding sites for cellular factors. Previous studies have indicated that in the context of the avian sarcoma virus genome, precise deletion of both ASV dr1 elements results in a very low level of virus replication. This is characterized by a decreased cytoplasmic accumulation of unspliced RNA and a selective increase in spliced src mRNA. Deletion of either the upstream or downstream dr1 results in a delayed-replication phenotype. To determine if the same regions of the dr1 mediate inhibition of src splicing and unspliced RNA transport, point mutations in the upstream and downstream elements were studied. In the context of viral genomes with single dr1 elements, the effects of the mutations on virus replication and increases in src splicing closely paralleled the effects of the mutations on CTE activity. For mutants strongly affecting CTE activity and splicing, unspliced RNA but not spliced RNA turned over in the nucleus more rapidly than wild-type RNA. In the context of wild-type virus containing two dr1 elements, mutations of either element that strongly affect CTE activity caused a marked delay of virus replication and a selective increase in src splicing. However, the turnover of the mutant unspliced RNA as well as the spliced mRNA species did not differ significantly from that of the wild type. These results suggest the dr1 elements in ASV act to selectively inhibit src splicing and that both elements contribute to the fitness of the wild-type virus. However, a single dr1 element is sufficient to stabilize unspliced RNA.

Animals↗
PubMed2001

A new RNA element located in the coding region of a murine endogenous retrovirus can functionally replace the Rev/Rev-responsive element system in human immunodeficiency virus type 1 Gag expression.

Nuclear export of incompletely spliced RNAs is a prerequisite for retroviral replication. Complex retroviruses like human immunodeficiency virus (HIV) encode a viral transport factor (Rev), which binds to its target sequence on the RNA genome and directs it into the Crm-1-mediated export pathway. Other retroviruses, like Mason-Pfizer monkey virus, contain cis-acting constitutive RNA transport elements (CTE) which achieve nuclear export of intron-containing RNA via cellular transport factors. Here, we describe the identification and characterization of a novel cis-acting orientation-dependent RNA expression element in the coding region of the murine intracisternal A-type particle (IAP) MIA14. This IAP expression element (IAPE) can functionally replace the Rev system in the expression of HIV-1 Gag proteins but functions independently of Crm-1. The presence of this element is needed for the expression of the IAP Gag proteins, indicating its biological significance. The IAPE can be functionally replaced by placing a CTE on the MIA14 RNA, further supporting its role in mRNA export. Northern blot analysis revealed that total RNA, as well as cytoplasmic RNA, was increased when the element was present. The element was mapped to a predicted stem-loop structure in the 3' part of the pol open reading frame. There was no overall homology between the IAPE and the CTE, but there was complete sequence identity between short putative single-stranded loops. Deletion of these loops from the IAPE severely reduced Rev-independent Gag expression.

Animals↗
PubMed1999

Identification of novel import and export signals of human TAP, the protein that binds to the constitutive transport element of the type D retrovirus mRNAs.

The nuclear export of the unspliced type D retrovirus mRNA depends on the cis-acting constitutive transport RNA element (CTE) that has been shown to interact with the human TAP (hTAP) protein promoting the export of the CTE-containing mRNAs. We report here that hTAP is a 619-amino-acid protein extending the previously identified protein by another 60 residues at the N terminus and that hTAP shares high homology with the predicted rat and mouse TAP proteins. We found that hTAP is a nuclear protein that accumulates in the nuclear rim and the nucleoplasm. We further demonstrated that hTAP is able to shuttle between the nucleus and the cytoplasm. Identification of the signals responsible for nuclear import (NLS) and export (NES) revealed that they are distinct but partially overlapping. NLS and NES of hTAP are active transferable signals that do not share similarities with known elements. The C-terminal portion contributes further to hTAP's nuclear retention and contains a signal(s) for nuclear rim association. Taken together, our data show that hTAP is a dynamic protein capable of bidirectional trafficking across the nuclear envelope. These data further support hTAP's role as an export factor of the CTE-containing mRNAs.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed2005

Hemorrhage following coblation tonsillectomy.

OBJECTIVES: We performed a prospective study to evaluate the incidence of post-tonsillectomy hemorrhage (PTH) in adults and children who underwent Coblation tonsillectomy (CTE) under general anesthesia. METHODS: The data of 63 adults and children (mean age, 21.8 years) were analyzed. RESULTS: There were 7 episodes of considerable bleeding (11.1%) that required surgical treatment under general anesthesia in 6 patients, of whom 5 experienced secondary bleeding (>24 hours). Moreover, bleeding and massive swelling of the pharynx required surgical treatment and prolonged intubation (35 hours) in 1 patient. None of the patients received blood transfusions. There was no case with a lethal outcome. Less intense bleeding (clots; blood-tinged sputum) was observed in 17 patients (27%) who required readmission or prolonged inpatient observation, 1 of whom had previously undergone surgical treatment of PTH. However, these 17 patients had an uneventful clinical course. In total, 22 patients experienced minor or major forms of PTH (34.9%). CONCLUSIONS: At least in our hands, CTE dramatically increased the frequency of PTH. The high rate of secondary bleeding contrasts with our documented experience using conventional methods, ie, cold dissection and suture ligation, to achieve hemostasis (7.9% with CTE versus <0.8% with conventional methods). Therefore, at our institution, tonsillectomy with conventional instruments remains the method of choice.

Adult↗
PubMed2004

Center wavelength shift dependence on substrate coefficient of thermal expansion for optical thin-film interference filters deposited by ion-beam sputtering.

Single-layer films of Ta2O5 and multilayer thin-film filters of Ta2O5 and SiO2 were deposited by ion-beam-sputter deposition. Postdeposition annealing of the structures resulted in increased optical thickness of the films, resulting in an upward shift in the wavelength of the transmission-reflection spectra. Modeling of the single-layer films by means of the effective media approximation indicates an increase in the void fraction of the film after annealing. This increase is consistent with an observed decrease in refractive index and an increase in physical film thickness. The multilayer structures, deposited on substrates of varying coefficient of thermal expansion (CTE), were annealed at various temperatures, and the change in the center wavelength was measured. The measured change is dependent on the annealing temperature and the substrate CTE, indicating that the increase in the void fraction is caused in part by thermally induced stress during the annealing process. A simple model is proposed that relates the void fraction present in the films after annealing with the substrate CTE and the annealing temperature.

Journal Article↗
PubMedDate not supplied

Crossed testicular ectopia.

Crossed testicular ectopia (CTE) is a rare anomaly, characterized by migration of one testis towards the opposite inguinal canal. Presented here is a case of crossed ectopia of the right testis, treated by extraperitoneal transposition of the gonad and right orchiopexy. Embriology and surgical findings suggest that CTE is a common consequence of many unclear ethiologic factors, specially mechanical ones, and can be associated with Muller duct persistence. Review of literature suggests a classification of CTE into 3 types: I--associated with inguinal hernia alone; II--associated with persistent mullerian remnants; III--associated with other anomalies without mullerian remnants. Treatment includes transeptal orchiopexy or extraperitoneal transposition of the testis, research for mullerian remnants and other anomalies, and long term postoperative follow-up, due to the risk of becoming malignant.

Child, Preschool↗
PubMed1975

[The effect of diphtheria toxin on proliferation of a transplantable culture of human amnion cells].

Cytoxic effect (CTE) of diphtheria toxin (DT) is preceded by disturbances of proliferation process, this being expressed in reduction of the proliferation index (PI) and of mitotic index (MI) before the appearance of the CTE. The effect of the toxin on the proliferation and mitoses of the cell culture is expressed after the incubation period of about 3 hours. The extent and the rate of reduction of the indices directly depended on the dose of the toxin whose low doses at the early periods stimulated the proliferation. The maximum of all the manifestations of the toxin action coincided with the period of increased proliferation in control. On the basis of investigations carried out a hypothesis was put forward that the CTE of diphtheria toxin developed with the entrance of the cells into mitosis.

Amnion↗
PubMed2001

[Cytotoxic effect of Acanthamoeba trophozoite on HeLa cells].

OBJECTIVE: To investigate the cytotoxic effect(CTE) on human cervix cancer HeLa cells induced by five strains of pathogenic free-living Acanthamoeba. METHODS: The cytotoxic effect of five isolates of Acanthamoeba on HeLa cells was investigated by light microscopy and MTT method. RESULTS: The photomicrographs of HeLa cells showed a sequence of cardinal morphological features of apoptosis when HeLa cells were exposed to Acanthamoeba in a time-dependent manner at a ratio of 1:1 for 12 h. MTT method showed more than 50% of tumor cells underwent cytolysis following exposure to A. lugdunensis trophozoites, and only 18% of cells treated with A. polyphaga underwent CTE. The CTE produced by A. lugdunensis and A. quina trophozoites was more rapid than the others, beginning as early as 6 h after coincubation and resulting in cytolysis by 72 h. CONCLUSION: These five strains of Acanthamoeba exhibit cytotoxic effects of varying degrees on HeLa cells, inducing apoptosis.

Acanthamoeba↗
PubMed2003

[Local application of bFGF and sucralfate during continuous tissue expansion].

OBJECTIVE: To investigate the effect of local application of bFGF and sucralfate during continuous tissue expansion (CTE). METHODS: CTE combined with local administration of bFGF and sucralfate was used in twelve patients with scar and nasal tip defects. Twenty three expanders were placed in the subcutaneous pockets through intralesion short incisions. Continuous expansion began at 1-3 days after expander implantation. The histomorphological changes and epidermal cell proliferation were observed. The clinical results were investigated. RESULTS: The average inflation time was 8.9 days. The average interval of the two operations was 13.5 days. The average hospitalization was 28.4 days. The average immediate stretch-back rate of the expanded skin was 25.7%. The clinical results were satisfactory without any complications. Histological examinations showed that the epidermal, granular and spinous layer became thicker. The basal cells increased significantly. The dermis thinned slightly and the collagen fibers became thicker. The elastic fiber regenerated significantly. Fibroblast and capillary density increased obviously. The immunohistochemistry analysis showed that the proliferation of epidemic basal cells was significant postoperatively. CONCLUSION: Local application of exogenous bFGF and sucralfate during CTE was feasible in patients. It could accelerate tissue expansion and improve the quality of expanded skin flap.

Capillaries↗
PubMed1990

A comparative study on reductive dehalogenation of halothane in liver, kidney and lung of the rabbit.

The contents of cytochrome P-450 (P-450) and cytochrome b5, and the activity of NADPH-cytochrome c reductase and the reductive metabolites of halothane, 2-chloro-1, 1-difluoroethylene (CDE) and 2-chloro-1, 1, 1-trifluoroethane (CTE) were measured in microsomes from the liver, kidney and lung of phenobarbital (PB) pretreated and untreated Japanese white strain rabbits. Microsomal P-450 levels in the liver, kidney (renal cortex) and lung of the rabbits were 1.91 +/- 0.35, 0.19 +/- 0.04 and 0.42 +/- 0.11 nmol/mg protein (mean +/- SD), respectively. In vivo phenobarbital pretreatment (PB-pretreatment) increased the content of P-450 to 2.95 +/- 0.40 nmol/mg protein (154%) in the liver and to 0.40 +/- 0.11 nmol/mg protein (211%) in the kidney, but had little effect in the lung. The activity of CDE formation was 0.72 +/- 0.10, 0.08 +/- 0.04 and 0.03 +/- 0.01 nmol/mg protein/min in the liver, kidney and lung, respectively. PB-pretreatment enhanced the activity of CDE formation to 1.59 +/- 0.49 nmol/mg protein (221%) in the liver, and to 0.29 +/- 0.16 nmol/mg protein/min (363%) in the kidney, but showed little enhancement in the lung. The activity of CTE formation was 1.30 +/- 0.19, 0.12 +/- 0.04 and 0.09 +/- 0.02 nmol/mg protein/min, in the liver, kidney and lung, respectively. PB-pretreatment enhanced the activity of CTE formation to 1.80 +/- 0.44 nmol/mg protein/min (138%) in the liver, but caused only slight enhancement in the kidney and lung. PB-pretreatment markedly enhanced the activity of CDE formation in the kidney. The authors conclude that cytotoxicity by reductive dehalogenation of halothane is possible not only in the liver but also in the kidney with PB-pretreatment.

Animals↗
PubMed1990

[Image processing of topographic EEG in epilepsy].

The processing of epileptic computerized topographical EEG (CTE) with the principle of image enhancement and point-by-point process was reported. The results indicated that the high potential distribution of CTE markedly reduced, the focus location was prominent and the mirror nearly disappeared after processing CTE image. The difference increased between the spike grey and the fast wave. This is advantageous to the surgical treatment of epileptic focus in brain.

Adolescent↗
PubMed1990

A specific cis-acting element regulates in vitro transcription of sex-dependent mouse steroid 16 alpha-hydroxylase (C-P450(16 alpha)) gene.

The mouse steroid 16 alpha-hydroxylase (C-P450(16 alpha)) family consists of structurally similar genes with very divergent expressions (Wong, G., Itakura, T., Kawajiri, K., Skow, L., and Negishi, M. (1989) J. Biol. Chem. 264, 2920-2927). Of five family members, the C-P450(16 alpha) gene exhibits a male-specific expression whereas the P450cb gene is equally expressed in both males and females although the level of expression is much lower than that of the C-P450(16 alpha) gene. By DNase I footprinting and in vitro transcription of the C-P450(16 alpha) gene, we determined two cis-acting transcription elements. SDI (sex difference information) in the -84/-102 region from the transcription start site is one of the two cis-acting transcription elements, which is specific to the sex-specific C-P450(16 alpha) gene and confers a high transcription level of this gene. We substituted the nucleotide(s) in the SDI sequence to the corresponding base(s) in the P450cb gene promoter and tested each substituted gene promoter by in vitro transcription. The results indicate that the function of SDI as the transcription element depends most critically on the nucleotides at positions -100 to -98. Another cis-acting transcription element, CTE (common transcription element), is located between -44 and -68 in both C-P450(16 alpha) and P450cb genes. A column chromatography of nuclear extract indicates that the proteins bound to SDI and CTE are different. We speculate therefore that SDI is a specific and necessary element involved in the male-specific C-P450(16 alpha) gene transcription. The sequence comparisons of the corresponding regions of SDI and CTE among the mouse and the closely related rat P450 genes suggest that SDI is a newly evolved cis-acting element in the sex-specific C-P450(16 alpha) gene which arose through gene duplication and selective nucleotide substitution.

Animals↗
PubMed1986

Production of enterotoxins by strains of Escherichia coli isolated from pigs.

The production of enterotoxins by 237 hemolytic strains of Escherichia coli isolated from pigs was determined with the use of CTE in CHO. Vero and Hela cells and ILT. More frequent (p less than 0.01) production of enterotoxins, determined by ILT, was found for the serotypes being pathogenic for the animals (63.8% of the strains). No correlation between intensity of ILT and particular serotype was observed. Both the serotypes pathogenic for pigs and other serotypes produced LT enterotoxins and ST toxin. The frequency of LT enterotoxin production was statistically insignificant compared to the frequency of ST enterotoxin production by strains with serotypes pathogenic for the pigs. Strains of E. coli producing only enterotoxin ST belonged both to the pathogenic serotypes as well as to other hemolytic serotypes. The cytotoxic activity of supernatants of E. coli strains with different serotypes isolated from pigs in Vero and Hela cells and simultaneous CTE in CHO cells was observed. This suggests the production by the strains of enterotoxin LT and cytotoxin VT. Seven out of the 96 isolates showing CTE in CHO cells gave no reaction in the ILT in pigs. This suggests the production by these isolates of a toxin (toxins) differing from the E. coli enterotoxins.

Animals↗
PubMed1989

Macrophage-lymphocyte interaction in the formation of delayed-type hypersensitivity.

Macrophage-lymphocyte interaction was studied on 121 CBA mice during a 2-hour contact of lymph-node cells of non-immune mice with a monolayer of peritoneal macrophages of BCG-immunized mice and subsequent intravenous administration of 4.10(7) pre-incubated lymphocytes to syngenic recipients. Sensitivity to tuberculin was demonstrated in the recipients by means of blast-transformation reaction of spleen cells (stimulation index was evaluated according to incorporation of 3H-thymidine--SI = 1.32 +/- 0.40) using administration of tuberculin into the paws (Mantoux reaction--MR = 0.11 +/- 0.02 mm) and the cytotoxic effect (CTE) of the lymphocytes on tuberculin-loaded sheep-cell erythrocytes whose disintegration was assessed according to discharge of iron from the target cells (CTE = 13.98 +/- 2.73%). At transfer of intact lymphocytes after contact with non-immune macrophages, SI = 1.046 +/- 0.019; MR = 0.014 +/- 0.002 mm; CTE = 0.214 +/- 0.048%. The treatment of lymphocytes with indomethacin during the contact with macrophages induced idvere changes in the indices of delayed-type hypersensitivity (DTHS). The conclusion has been drawn that the antigen-presenting ability of macrophages can materialize in vitro.

Animals↗
PubMedDate not supplied

Metabolic activation of the halothane metabolite, [14C]2-chloro-1,1-difluoroethene, in hepatic microsomes.

Halothane is reduced to 2-chloro-1,1,1-trifluoroethane (CTE) and 2-chloro-1,1-difluoroethene (CDE) by cytochrome P-450. These compounds may potentially undergo secondary metabolism in vivo, but their capacity to undergo metabolic activation and bind to macromolecules is unknown. This study, therefore, compared the abilities of CDE and CTE to bind to microsomal components in relation to that of halothane in hepatic microsomes. The results show that CDE, in addition to halothane, binds to microsomes under conditions of cytochrome P-450 activity. While halothane bound predominantly to lipids under nitrogen, CDE bound mainly to protein under oxygen. No CTE binding under any conditions could be detected. On an equimolar basis, CDE binding to protein was approximately one-third of that of halothane under oxidative conditions, however, CDE binding was enhanced in the presence of halothane. The results support the hypothesis that CDE metabolism may contribute to the metabolic binding due to halothane exposures.

Anaerobiosis↗
PubMedDate not supplied

[Triiodothyronine thyrotoxicosis in nodular toxic goiter].

Serum protein-bound iodine (PBI), total blood thyrotoxin (T4), triiodthyronine (T3), coefficient of thyrotoxin efficacy (CTE), serum thyrotoxin-binding capacity (TBC) or some of these indices were determined in 44 patients with nodular toxic goiter with thyrotoxemia. An increase in T3 level was revealed in 27 of 35 of the patients; T3-thyrotoxicosis was diagnosed in 18 of these in whom an increase of the T3 level was combined with normal T4 or normal CTE content and an unchanged TBC. The symptoms of T3-thyrotoxicosis displayed no significant differences from the symptoms of thyrotoxicosis caused by the excess of thyroxin. In many of the patients T3-thyrotoxicosis coursed for a long time with but a few symptoms. Loss of weight in this form of thyrotoxicosis was much less frequent than in "thyrotoxin'' thyrotoxicosis. PBI was elevated in 12 of 44 patients, and total T4 and CTE -- in 10 of 35. These methods do not permit to diagnose hyperthyroidism in many of the patients with nodular toxic goiter in which T3-thyrotoxicosis is of frequent occurrence.

Adult↗
PubMedDate not supplied

Inhibitory effect of gomisi on reductive metabolism of halothane.

The effect of Gomisi (dried ripe fruit of schizandra chinensis) on chlorodifluoroethylene (CDE) and chlorotrifluoroethane (CTE) formation was investigated. The incubation mixtures for the measurement of reductive metabolites of halothane consisted of liver microsomal suspensions, 3 mM NADPH, extract solution of Gomisi and halothane in 0.1 M potassium phosphate buffer (pH 7.4). The production of CDE and CTE was inhibited by Gomisi in a dose-dependent way. The production were reduced to half in the presence of 0.5% Gomisi extract in the reaction mixture. The results suggest that Gomisi can inhibit the reductive metabolism of halothane in vitro; thus it may protect against halothane-induced hepatitis.

Animals↗
PubMed2001

Evaluation of potential neurotoxic effects of occupational exposure to (L)-lactates.

Organo psycho syndrome (OPS) or chronic toxic encephalopathy (CTE) is a neurotoxic condition reported following long-term exposure to paints containing organic solvent and to other solvents. Lactate esters are finding wider use as solvents. Lactate esters have been well studied in standard toxicity tests, but specific neurotoxicity studies have not been conducted. No clinical signs of chronic neurotoxicity have been observed in standard toxicity tests. Lactate esters are rapidly hydrolyzed in the body to lactic acid and the corresponding alcohol. Alcohols have been reported to have acute neurotoxic effects, usually following high levels of ingestion. The literature on alcohols was reviewed to establish the no-observed-adverse-effect level (NOAEL) for acute neurotoxicity and to look for any evidence of chronic neurotoxicity from the alcohols produced by hydrolysis of the lactate esters. The NOAELs were compared with the potential amounts of alcohol produced by hydrolysis of different lactate esters at 200 mg//m(3) (the NOAEL for most of the lactate esters). In all cases neither acute nor chronic neurotoxicity would be expected based on the amounts of alcohol produced by hydrolysis of the lactate esters at their NOAELs. L-Lactic acid is a normal metabolite in the body and is not considered neurotoxic. Based on this information there is no evidence to suggest that L-lactate esters can cause any chronic neurotoxicity, OPS, or CTE.

Alcohols↗