Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Five microwell non isotopic hybridization assays, based on colorimetric immunoenzymatic reading, were developed and evaluated for the rapid and automatable detection of enteroviruses. Virus nucleic acids and/or capture probes were covalently bound to microtiter wells, and digoxigenin-11-dUTP was used as label for the detection of hybridized material. Among these procedures, a reverse transcriptase polymerase chain reaction (RT-PCR) hybridization assay was the most sensitive, enabling the detection of 10 MPNCU of poliovirus, and offering detection specificity for other enteroviruses, such as coxsackieviruses and echoviruses. The second most sensitive method was a complementary hybridization assay, simultaneously using three detection probes, one from the 5' end and two from the 3' end of poliovirus genome, offering a sensitivity for poliovirus detection of 5 x 10(3) MPNCU.
Mol Cell Probes 1996 Apr
PMID:Non isotopic automatable molecular procedures for the detection of enteroviruses. 873 91

Apoptosis, a highly regulated and energy-dependent process of cell death, plays an important part in normal tissue development. Its role in pathological conditions may vary. Earlier morphologic studies suggest that apoptosis may be an important mechanism in light-induced photoreceptor degeneration. In this study, we determined if photic exposure triggers apoptosis in photoreceptor cells in an established model of photoreceptor degeneration by light exposure. Twenty eight Lewis albino rats were divided into seven groups of 4 animals each: one group served as the unexposed control; and the other groups were exposed continuously to green fluorescent light (320 foot-candles) for 3, 6, 9, 12, 18 or 24 hours, respectively and killed for histopathological examination, biochemical isolation of retinal DNA; and in situ analysis of nicked nuclear DNA by terminal transferase biotin-dUTP nick end labeling (TUNEL). Histopathological study revealed morphological changes comparable to previous reports on photic injury. Electrophoretic analysis of DNA showed internucleosomal DNA cleavage as early as 12 hours of light exposure. The fluorescent intensity of DNA fragments, which were monomers and multimers of 180-200 base pairs, increased with the duration of light exposure. TUNEL technique, which localized DNA cleavage to the photoreceptor cell nuclei as early as 6 hours of light exposure, showed that the number of TUNEL positive nuclei increased with light exposure, and revealed more DNA degradation in the superior quadrant. Our findings confirmed earlier morphologic observations that photic exposure triggers apoptosis of photoreceptor cells.
Res Commun Mol Pathol Pharmacol 1996 May
PMID:Photic injury triggers apoptosis of photoreceptor cells. 877 71

Strand Displacement Amplification (SDA) is an isothermal, in vitro method of amplifying DNA that is based upon the combined action of a DNA polymerase and restriction enzyme. Previously, a form of SDA was developed which utilizes the exonuclease deficient Klenow fragment of E. coli polymerase I (exo Klenow) and the restriction enzyme HincII to achieve 10(8)-fold amplification in 2 h at 37 degrees C (Walker, G.T., 1993, PCR Methods and Applications 3; 1-6). A new thermophilic form of SDA is reported here which uses a restriction endonuclease from Bacillus stearothermophilus (BsoBI) and a 5'-->3' exonuclease deficient polymerase from Bacillus caldotenax (exo Bca). SDA was used to amplify DNA from Mycobacterium tuberculosis. An amplification factor of 10(10)-fold was achieved after 15 min of SDA at 60 degrees C. The new thermophilic system is much more specific than the previous mesophilic system as evidenced by a dramatic decrease in background amplification products. Thermophilic SDA was also optimized with dUTP substituted for TTP to enable amplicon decontamination using uracil-DNA glycosylase.
Mol Cell Probes 1996 Aug
PMID:Detection of M. tuberculosis DNA using thermophilic strand displacement amplification. 886 73

An ovariole-specific yolk polypeptide cDNA from Galleria mellonella was isolated from a cDNA library constructed using vitellogenic ovariole poly (A)+ RNA, by differential screening with the ovariole and day-0 male pupal poly (A)+ RNA. The Ov11 cDNA hybridized to a 1.1 kb message from the ovariole but not from male pharate adults, ovariectomized pharate adults or day-5 last instar larvae. Developmental Northern analysis showed that this message is expressed only during vitellogenic stages. In situ hybridization of digoxigenin-dUTP labeled Ov11 cDNA to the day-8 pharate adult ovariole showed that only follicle cells express the Ov11 gene. The 960 bp Ov11 cDNA is nearly full-length, containing a single open reading frame coding for a 286 amino acid polypeptide with a hydrophobic signal sequence and two potential N-glycosylation and tyrosine sulfation sites. The fact that Ov11 cDNA codes for the 37 kDa ovariole-specific yolk protein, YP4, was confirmed by the N-terminal amino acid sequence analysis of purified YP4. Genomic Southern analysis suggests that the yp4 gene is a single copy gene and PCR analysis using the 5' and 3' end primers of the cDNA indicates that the yp4 gene is intronless within the coding region.
Insect Biochem Mol Biol 1996 Jun
PMID:Isolation, characterization and complete nucleotide sequence of a Galleria mellonella cDNA coding for the follicle cell-specific yolk protein, YP4. 896 66

The incidence of apoptosis and the expression of Fas antigen (Fas)/Fas ligand (FasL) mRNA in bleomycin-induced pulmonary fibrosis in mice were examined. Male ICR mice were intratracheally instilled with bleomycin (5 U/kg of body weight). The controls were injected with sterile saline. The animals were anesthetized and killed at 1, 6, and 12 h, and 1, 3, 5, 7, 9, and 14 days after bleomycin instillation. We assessed the incidence of apoptosis in lung tissues by DNA fragmentation on agarose gel electrophoresis, terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end-labeling, and electron microscopy. The expression of Fas and FasL mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). The localization of Fas mRNA was analyzed by in situ hybridization and that of FasL mRNA was analyzed by RT in situ PCR. The results showed that (1) a single instillation of bleomycin leads to the rapid appearance of apoptosis in bronchial and alveolar epithelial cells, which resolves within 1 day, and (2) apoptosis reappears on day 7 and continues for over 14 days after bleomycin instillation. This was accompanied with a progression of fibrosis. Corticosteroid administration completely blocked both apoptosis and fibrosis. The expression of Fas mRNA was upregulated in the alveolar epithelial cells by the bleomycin instillation. FasL mRNA was also upregulated in infiltrating lymphocytes after bleomycin treatment, but not in the control mice. The administration of corticosteroids suppressed the expression of Fas and FasL mRNA as well as apoptosis and fibrosis. Although these results do not show that apoptosis mediated by the Fas/FasL system is directly linked to bleomycin-induced fibrosis, we speculate that excessive apoptosis and the Fas/FasL system play a role in the pathogenesis of bleomycin-induced lung injury.
Am J Respir Cell Mol Biol 1997 Jan
PMID:Apoptosis and expression of Fas/Fas ligand mRNA in bleomycin-induced pulmonary fibrosis in mice. 899 84

Occurrence of Pseudomonas aeruginosa, Stenotrophomonas (Xanthomonas) maltophilia and Burkholderia (Pseudomonas) cepacia in sputum of cystic fibrosis (CF) patients was demonstrated with a simple and rapid polymerase chain reaction (PCR) technique. The PCR was performed with a set of three primer pairs based on 16S rRNA sequences after sputum preparation with dithiothreitol and NaOH lysis. All three pathogens could be individually detected by the use of this technique. To prevent carry-over contamination, dUTP and uracil-N-glycosylase were included in the reaction. The amplicons were visualized by agarose gel electrophoresis. Sputum culture was performed on all samples. Ninety specimens from CF patients were analysed. The sensitivity for the detection of P. aeruginosa was 37/40 (93%) compared to culture. Bacterial growth of P. aeruginosa was found in three cases, where PCR amplicons were not detected, while PCR was positive in five cases, where culture did not reveal the presence of this bacterium. For this reason, the specificity was 45/50 (90%). For S. maltophilia, the PCR was less sensitive than culture (positive in three of six cases). In our series, B. cepacia was detected by culture in one case and this was also detected by PCR. There were no false-positive PCR results regarding S. maltophilia or B. cepacia. Thus, combined PCR-based detection of these three clinically relevant bacteria in sputum samples from CF patients can be performed by a reliable technique in a relatively simple manner. The present data indicate a high sensitivity and specificity for P. aeruginosa. The lower sensitivity observed for the detection of S. maltophilia in sputum and B. cepacia, as estimated from laboratory strains, may depend on PCR conditions and genetic heterogeneity, respectively. The greatest gains with this method can be made when it is used for the early detection of P. aeruginosa in sputum-producing CF patients.
Mol Cell Probes 1996 Dec
PMID:Polymerase chain reaction for the detection of Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Burkholderia cepacia in sputum of patients with cystic fibrosis. 902 76

This study was carried out to develop a rapid and simultaneous detection system of chromosome Y- and 1-bearing porcine spermatozoa by fluorescence in situ hybridization (FISH). Chromosome Y- and 1-specific DNA probes were produced by polymerase chain reaction with digoxigenin (Dig)- or biotin-dUTP. The hybridization probe mixture of labeled Y-chromosome and chromosome 1-specific DNA was applied to the preparation, immediately denatured at 75 degrees C for 8 min, hybridized for 5 min at 37 degrees C and overall FISH steps were done within a few hours. When double FISH with Dig-labeled chromosome Y-specific and biotin-labeled chromosome 1-specific probes was applied to sperm nuclei pretreated with dithiothreitol, the average of 50.9% of sperm nuclei had the Dig-signal, 99.2% of the sperm nuclei had the biotin-signal and the average of 0.3% of sperm nuclei showed no signal. The putative rate of Y-bearing spermatozoa ranged from 49.8% to 52.8% among 5 boars and the average putative rate of Y-bearing spermatozoa was 51.0%. The results indicated that a rapid and simultaneous FISH with chromosome Y- and 1-specific porcine DNA probes produced by PCR made possible more accurate assessment of Y-bearing porcine spermatozoa.
Mol Reprod Dev 1996 Apr
PMID:Rapid and simultaneous detection of chromosome Y- and 1-bearing porcine spermatozoa by fluorescence in situ hybridization. 905 47

This study investigated terminal dUTP nick-end labeling (TUNEL)-positive cells in the frontal, occipital, and hippocampal cortices of seven normal aging and four Alzheimer's patients. Significant increase in TUNEL-positive cells was observed in the frontal and hippocampal cortices of Alzheimer's patients when compared with controls. In the hippocampal cortex, only area CA4 demonstrated a significant increase of TUNEL-positive cells. Double staining of TUNEL-positive cells for glial fibrillary acidic protein revealed that < 13% of the TUNEL-positive nuclei belonged to astrocytes. The results of this study illustrated a differential pattern of cortical degeneration between normal aging and Alzheimer patients.
J Mol Neurosci 1997 Apr
PMID:Terminal dUTP nick end labeling (TUNEL) positive cells in the different regions of the brain in normal aging and Alzheimer patients. 918 38

To clarify whether apoptosis is involved in endometriosis, we obtained eutopic endometrial tissues along with endometriotic tissues from the uterus (adenomyosis) (n = 12) and from the ovary (n = 12) from patients undergoing gynaecological surgery. Apoptosis-induced DNA fragmentation was detected by the TdT-mediated dUTP-biotin nick-end labelling method, and immunostaining with a monoclonal antibody against the Fas, Le(y) or B-cell leukaemia/lymphoma-2 (bcl-2) was also performed using the same tissue section. Analysis showed that apoptosis was occurring in all the samples of ovarian endometriotic tissue but in only two of the 12 adenomyotic and in five of the 24 eutopic endometrial tissue samples. In none of these cases was apoptosis correlated with phases of the menstrual cycle. The expression of bcl-2 in the eutopic endometrial and adenomyotic tissues was limited to the proliferative phase, and was observed in only one of the 12 cases of ovarian endometriosis. Fas and Ley were expressed randomly across a wide range in both the eutopic and ectopic endometrial tissues. These results suggest that the features of ovarian endometriosis are different from those of adenomyosis and eutopic endometrium in terms of the involvement of apoptosis. In addition, the regulatory mechanism involved in ovarian endometriosis may differ from that in other endometrial cells.
Mol Hum Reprod 1996 May
PMID:Detection of apoptosis in human endometriotic tissues. 923 97

The purpose of this study was to document induction of apoptosis by vitamin E succinate (VES; RRR-alpha-tocopheryl succinate) in human breast cancer cells in culture and to characterize potential c-jun involvement. VES at 18.8 microM (10 micrograms/mL) induced DNA synthesis arrest, reduced total cell numbers, and induced apoptosis in estrogen receptor-positive and estrogen-responsive MCF-7 human breast cancer cells. VES at 10 micrograms/mL induced apoptosis in greater than 60% of cells within 3 d of treatment. Apoptosis was documented by detection of fragmented or condensed nuclei in 4',6-diamindino-2-phenylindole-stained cells, detection of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeled DNA, and DNA laddering. Analyses of mRNA and protein levels of candidate molecules involved in apoptosis showed that MCF-7 cells treated with VES exhibited elevated and persistent expression of c-jun. MCF-7 cells stably transfected with a dominant-negative interfering mutant c-jun, TAM-67, and expressing high levels of mutant jun exhibited approximately 50% blockage of VES-mediated apoptosis. In addition to increased c-jun expression after VES treatment, VES-treated MCF-7 cells exhibited elevated activator protein-1 (AP-1) binding activity. Comparisons of AP-1 binding factors by super-shift analyses with jun-specific antibodies in cells sensitive to VES-induced apoptosis (empty-vector control 7-1 cells) and cells resistant to VES-induced apoptosis (TAM-67-containing TAM-9 cells) showed that the sensitive cells expressed c-jun and jun D and the resistant cells TAM-67 AP-1 binding proteins after VES treatment. These studies suggested that c-jun may be involved in the apoptotic process initiated by VES treatment of human MCF-7 breast cancer cells.
Mol Carcinog 1997 Jul
PMID:Involvement of activator protein-1 (AP-1) in induction of apoptosis by vitamin E succinate in human breast cancer cells. 925 85


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>