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At least 973 records · Page 54Linked to original sources
PubMed1998

Quantitation of telomerase activity in hepatocellular carcinoma: a possible aid for a prediction of recurrent diseases in the remnant liver.

Because telomerase activity is necessary for cell immortality and probably associated with tumor progression, we have evaluated a possible aid for quantitation of the activity to predict intrahepatic recurrences after surgery in patients with hepatocellular carcinoma (HCC). HCC tissues obtained by surgical resection from 20 patients were studied. Telomerase activity was expressed as peaks with a periodicity through a fluorescence-based telomeric repeat amplification protocol using an autosequencer, and the quantity of activity was calculated from peak areas. A ratio of fluorescence intensity depending on telomerase to that of an internal standard was used as a value of relative telomerase activity (RTA). RTA in serially diluted S100 extracts from HepG2 cells was well correlated with the amount of the extracts. The mean RTA value of 36.4 +/- 27.8 (mean +/- SD, 3.21 to 105) in 9 patients suffering from early recurrences after surgery was significantly higher than that (9.84 +/- 7.65; mean +/- SD, 3.00 to 29.0) in 11 patients without intrahepatic recurrences during the early period (P = .004). These results indicate that RTA value can be a useful predictor for intrahepatic recurrences during the early period after surgical resection of HCC.

Adult↗
PubMed2006

Titanium implant osseointegration with calcium pyrophosphate in rabbits.

The objective of this study was to characterize calcium pyrophosphate material, evaluate its in vitro cytotoxicity, and assess its ability to induce bone formation. X-ray diffraction (XRD) was used to determine crystallinity and phases present in material. Serial dilutions of extracts, from 10-day dissolution tests in modified Eagle's medium, were exposed for 24 h to mouse fibroblasts and cytotoxicity assessed via viable staining. In vivo performance was determined by placing Ti screws with and without calcium pyrophosphate agglutinated with marrow adipose tissue in the tibiae of eight rabbits. New bone formation around test and control implants was evaluated histomorphometrically by using three fluorochrome labels: alizarin, calcein, and tetracycline. After 8 postoperative weeks, the animals were killed and specimens were retrieved and processed for fluorescence and light microscopic analysis. Calcium pyrophosphate showed no cytotoxicity and the XRD showed that the main phase of the analyzed sample corresponded to beta-calcium pyrophosphate. The largest fluorochrome labeling area occurred during the fourth and fifth postoperative weeks, in both control and experimental groups. Histologically, the bone neoformation occurred in regions where the calcium pyrophosphate was resorbed. The morphometric analysis showed implants placed with calcium pyrophosphate resulted in smaller polyfluorochrome labeling area (p < 0.05).

Animals↗
PubMed2001

Telomerase activity and telomere length in thyroid neoplasia: biological and clinical implications.

Despite several recent studies, the biological status and clinical relevance of telomerase expression in tumours derived from the thyroid follicular cell remain controversial. This study has analysed a series of normal, benign, and malignant thyroid samples using two novel approaches: the use of purified epithelial cell fractions to eliminate false-positives due to telomerase-positive infiltrating lymphocytes; and the simultaneous measurement of telomere length to provide a clearer interpretation of telomere dynamics in thyroid neoplasia. The data obtained support the prediction that the epithelial component of non-neoplastic thyroid and of follicular adenomas is telomerase-negative, any positive results being explicable by lymphocyte infiltration. In contrast, many malignant tumours, both follicular and papillary, were telomerase-positive. However, serial dilution of extracts indicated a wide spectrum of activity in these cancers, possibly related to variation in the proportion of telomerase-positive cells. Furthermore, an unexpectedly high proportion were telomerase-negative, a finding which was not explicable by technical problems such as TRAP (telomeric repeat amplification protocol) assay sensitivity. Many of these apparently telomerase-negative tumours had abnormally long telomeres. Correlation of telomerase and telomere length data suggests that thyroid cancers fall into three biological groups: telomerase-positive lesions, consistent with the conventional model of telomere erosion followed by telomerase reactivation; telomerase-negative tumours, which maintain telomere length by a mechanism independent of telomerase; and telomerase-negative tumours which are still undergoing telomere erosion and may therefore be composed of mortal cancer cells. From a clinical standpoint, it is concluded that telomerase detection on unfractionated tissue, such as fine needle aspirates, is of no value as a marker of malignancy in follicular lesions, due to both low sensitivity and specificity.

Adenoma↗
PubMed2002

Maturational changes in brain contents of salmon GnRH in rainbow trout as measured by a newly developed time-resolved fluoroimmunoassay.

A newly developed time-resolved fluoroimmunoassay (TR-FIA) for salmon gonadotropin-releasing hormone (sGnRH) was applied to investigate changes in sGnRH content in discrete brain areas at three different gonadal stages in the rainbow trout, Oncorhynchus mykiss. The sensitivity (6.8 pg/well), specificity, intraassay coefficients of variation (<7.4%), and interassay coefficients of variation (<10.3%) of the assay system were almost the same as those for the radioimmunoassay. Displacement curves of serially diluted brain extracts of nine teleost fish (freshwater fish and seawater fish) including rainbow trout paralleled that of the sGnRH standard, indicating that the sGnRH TR-FIA is widely applicable to the measurement of the brain sGnRH contents of various fishes. The sGnRH content in female hypothalamus decreased during final gonad maturation, whereas the sGnRH levels in pituitary were highest at the time of spermiating in males or ovulating in females, decreasing significantly thereafter. In contrast, there were no changes in the sGnRH contents of olfactory bulbs, telencephalon, optic tectum + thalamus, and cerebellum + medulla oblongata during final maturation, except for olfactory bulbs of males. Changes in sGnRH contents in the hypothalamus and the pituitary indicate that sGnRH is involved in final maturation (ovulation or spermiation) in the rainbow trout.

Aging↗
PubMed1981

Heterogeneous intrahepatic distribution of blood flow in humans.

Color-functional imaging of intrahepatic blood flow was developed using the 133Xe clearance method and a gamma camera with a computer system. During the 2 min after intrasplenic injection of 133Xe in saline solution, 24 sequential gamma images were obtained. After setting the hepatic region, 133Xe clearance curves were extracted from the serial images every 6 X 6 mm element and regional blood flow for each element was calculated. The calculated regional hepatic blood flow values were displayed as a color images in eight color steps. Eleven patients with and without liver diseases were studied. In all the patients studied, heterogeneous intrahepatic distribution of blood flow was clearly demonstrated by the functional image of regional hepatic blood flow. Although a consistent pattern of intrahepatic distribution of blood flow was not obtained, greater-flow regions were frequently observed in the right lobe. Repeat studies in two patients demonstrated that the intrahepatic distribution of blood flow varied. The 10-15 s scintiphotosplenoportograms also showed the existence of restricted or preferential intrahepatic distribution of splenic flow. These results strongly suggest that intrahepatic distribution of blood flow in the human liver is heterogeneous and variable.

Hepatitis↗
PubMed1991

Antigenic differences between Trichinella spiralis and T. pseudospiralis detected by monoclonal antibodies.

Antigenic differences between Trichinella spiralis and T. pseudospiralis were established using two monoclonal antibodies (mAbs) that show different specificities to muscle larvae of the two variants. Enzyme-linked immunosorbent assay (ELISA) revealed that mAb 3G6 reacts positively against T. spiralis, T. nelsoni, T. nativa and T. pseudospiralis, whereas mAb 3E10 does not react with T. pseudospiralis under the same experimental conditions. These antigenic differences were confirmed after preabsorption of the antibodies with serial dilutions of extracts of T. spiralis or T. pseudospiralis muscle larvae. The indirect immunofluorescence technique showed that the antigen corresponding to mAb 3G6 is located in the stichosomes and the cuticle surface of both T. spiralis and T. pseudospiralis. In contrast, mAb 3E10 positively stained cryostat sections of T. spiralis, forming a dense reaction product on the surface of the whole larvae and the surrounding capsule. This antibody can be quite useful as a specific probe for distinguishing T. spiralis from T. pseudospiralis in taxonomic studies. Using an avidin-biotin system, we could prove that mAb 3G6 recognizes an excretory/secretory-type antigen.

Animals↗
PubMed1985

Immunoassayable adrenocorticotropin in peripheral organs: concentrations during early development.

Although many have identified immunoassayable adrenocorticotropin (ACTH) in sites outside the pituitary (brain, gastrointestinal tract), there is relatively little information regarding immunoassayable ACTH in other major peripheral organs. Several major peripheral organs (pancreas, liver, kidney, heart) of rats were found to contain variable amounts of immunoreactive (IR-) ACTH which appeared to be authentic IR-ACTH on the basis of parallelism to ACTH1-39 upon serial dilution of extracts and gel filtration chromatography. Concentrations of IR-ACTH in peripheral organs were also studied to determine if changes occur during early development. Concentrations of IR-ACTH did not show significant changes in liver and heart at various ages between 10 and 80 days, but IR-ACTH in pancreas and kidney (day 10 vs. 80) did show significant decrements with aging.

Adrenocorticotropic Hormone↗
PubMed1993

Antigenic signature analysis reflects differences among plant virus isolates.

Antigenic differences among cowpea severe mosaic virus (CPSMV) isolates were clearly reflected in signature analysis employing a panel of seven well-characterized, monoclonal antibodies. Separate binding curves were generated by reacting serial dilutions of extracts from infected plant tissue containing each antigen simultaneously with each antibody in the panel. An iterative procedure was used to align unknown CPSMV antigen concentrations from different antigen preparations to allow comparison of binding profiles from different assays. Signature analysis was shown to be highly useful for the elucidation of subtle antigenic differences among viral agents because it requires neither purified virus nor knowledge of virus concentration in sap from infected plants.

Antibodies, Monoclonal↗
PubMed2004

Photoallergic contact dermatitis from ketoprofen induced by drug-contaminated personal objects.

BACKGROUND: Photoallergic contact dermatitis from ketoprofen has been recognized since the mid-1980s. Skin reactions have been reported to continue weeks after discontinuation of ketoprofen. One reason for this could be residual ketoprofen in the skin, which has been shown in a skin biopsy specimen. OBJECTIVE: We sought to report on 3 cases of photoallergic contact dermatitis from ketoprofen in topical anti-inflammatory gels and on relapses of dermatitis appearing after use of ketoprofen-contaminated objects. METHODS: We patch and photopatch tested, with standard series, the anti-inflammatory gel, ketoprofen, and its ingredients in serial dilutions and extracts of personal objects. We performed chemical investigations of personal objects with thin-layer chromatography, high-pressure liquid chromatography, and gas chromatography-mass spectrometry. RESULTS: Photoallergy was demonstrated to ketoprofen, which was detected in personal objects. CONCLUSION: Relapses of photoallergic contact dermatitis in patients photoallergic to ketoprofen can be induced by ketoprofen-contaminated objects such as bandages and slippers.

Administration, Topical↗
PubMed2006

Using fMRI to decompose the neural processes underlying the Wisconsin Card Sorting Test.

The specific role of particular cerebral regions with regard to executive functions remains elusive. We conducted a functional magnetic resonance imaging (fMRI) study to segregate different network components underlying the Wisconsin Card Sorting Test (WCST), a test widely applied clinically to assess executive abilities. Three different test variants of the WCST, differing in task complexity (A > B > C), were contrasted with a high-level baseline condition (HLB). Cognitive subcomponents were extracted in a serial subtraction approach (A-C, A-B, B-C). Imaging data were further subjected to a correlational analysis with individual behavioral parameters. Contrasting A with the HLB revealed the entire neural network underlying WCST performance, including frontoparietal regions and the striatum. Further analysis showed that, within this network, right ventrolateral prefrontal cortex related to simple working memory operations, while right dorsolateral prefrontal cortex related to more complex/manipulative working memory operations. The rostral anterior cingulate cortex (ACC) and the temporoparietal junction bilaterally represented an attentional network for error detection. In contrast, activation of the caudal ACC and the right dorsolateral prefrontal cortex was associated with increased attentional control in the context of increasing demands of working memory and cognitive control. Non-frontal activations were found to be related to (uninstructed relative to instructed) set-shifting (cerebellum) and working memory representations (superior parietal cortex, retrosplenium). The data provide neural correlates for the different cognitive components involved in the WCST. They support a central role of the right dorsolateral prefrontal cortex in executive working memory operations and cognitive control functions but also suggest a functional dissociation of the rostral and caudal ACC in the implementation of attentional control.

Brain↗
PubMed2003

Comparison of polymerase chain reaction and cell culture for the detection of Chlamydophila species in the semen of bulls, buffalo-bulls, and rams.

Two hundred and thirty six semen samples were collected from 120 bulls, 60 buffalo-bulls, and 56 rams located on farms of known history of infection with Chlamydophila species. All semen samples were examined by polymerase chain reaction (PCR) and cell culture techniques for detection of Chlamydophila species. The primers were selected to allow the amplification of all target species in a single reaction by identifying conserved sequences in the omp2 gene. PCR assay detected more positive samples (36) from the semen samples collected from different animal species than were detected by the culture method (21). The results indicated that all culture-positive semen samples (21) from different species were PCR positive. The detection limit of the PCR assay was determined with DNA extracted from fourfold serial dilution of C. abortus (B577) and C. pecorum (11/88) cultures and found to be 0.25 inclusion-forming units (IFU) per PCR, while the culture method could not detect less than 4 IFU. This is the first report using PCR for the detection of Chlamydophila species in buffalo-bulls' semen and the assay provides a simple, sensitive, rapid, and reliable means for the detection and identification of the organism.

Animals↗
PubMed2001

Fluorous Boc ((F)Boc) carbamates: new amine protecting groups for use in fluorous synthesis.

The first fluorous variants of the Boc (tert-butyloxycarbonyl) group have been prepared and tested for their suitability as nitrogen protecting groups. A group with two fluorous chains and an ethylene spacer, (RfCH2CH2)2(CH3)COC(O)-, was readily attached to a representative amine but was difficult to cleave. In contrast, groups with two fluorous chains and a propylene spacer, (RfCH2CH2CH2)2(CH3)COC(O)-, or one fluorous chain and an ethylene spacer, (RfCH2CH2)(CH3)2COC(O)-, were readily formed and cleaved. The fluorous alcohol component of the (F)Boc group can be removed by evaporation and can be recovered and reused. The utility of the new (F)Boc group (C8F17CH2CH2)(CH3)2COC(O)- was demonstrated in 16 and 96 compound library synthesis exercises. Separations can be achieved either by manual, parallel fluorous solid-phase extraction, or automated, serial fluorous chromatography. The results provide additional confirmation of the value of "light" fluorous synthesis techniques, and the new fluorous Boc groups expand the applicability of fluorous synthesis techniques to many classes of nitrogen-containing organic compounds.

Amines↗
PubMed1967

A procedure for the quantitative analysis of the sulphur amino acids of rat tissues.

1. A method is described for the quantitative separation of the sulphur compounds in a single sample of tissue by passing an extract through a serial assembly of ion-exchange resins in the order: Dowex 2 (Cl(-) form), Dowex 1 (CO(3) (2-) form), Amberlite CG-50 (H(+) form) and Zeo-Karb 225 (H(+) form). 2. Groups of sulphur amino acids were eluted separately from each column; the recovery of sulphur compounds after their labelling with (35)S in vivo by injection of l-[(35)S]-methionine was 91-106%. Individual sulphur compounds were further resolved by one-dimensional or two-dimensional paper chromatography. 3. Evidence is presented on the occurrence of S-adenosylmethionine and S-adenosylhomocysteine in rat liver and brain. Rat liver and brain contained 83.6 and 31.4mmu-moles/g. respectively of S-adenosylmethionine.

Journal Article↗
PubMed2002

Studies of piroxicam absorption by oral mucosa.

The present study was undertaken to investigate, if the non-steroidal anti-inflammatory drug (NSAID) piroxicam (CAS 36322-90-4) Fast-Dissolving Dosage Form (FDDF) can be absorbed in the oral mucosa. Piroxicam FDDF was administrated under the tongue to rats with an oesophagus ligation (OL) to prevent the drug entering the stomach and in turn its absorption by the classic way. A group of sham-operated (SO) animals received the same piroxicam FDDF dose. After drug administration, a pharmacokinetic study with serial serum sample extractions at 0, 15, 30, 60 and 120 min was performed. It has been found a prompt increase in serum piroxicam levels in OL-rats, which showed different pharmacokinetics from SO-rats. Areas under curve (AUCs) of OL-rat serum piroxicam levels were higher at 15, 30 and 60 min compared to SO-animals. These results indicate that piroxicam FDDF is absorbed in the rat oral mucosa. Moreover, during the first hour, drug absorption by oral mucosa rendered higher piroxicam levels than gastric absorption.

Absorption↗
PubMed1982

Localization of peptide YY (PYY) in gastrointestinal endocrine cells and effects on intestinal blood flow and motility.

In immunohistochemical studies using antisera to peptide YY (PYY), a 36 amino acid polypeptide isolated from porcine duodenum, it was found that PYY-like immunoreactivity occurred mainly in endocrine cells of the gastrointestinal mucosa. PYY-immunoreactive cells were particularly abundant in the distal intestine and have been observed in five species, including man. By radioimmunoassay it was found that, in the rat, the amount of PYY immunoreactivity was about 100-fold higher in the colon than in the duodenum. The chromatographic profiles of PYY immunoreactivity from the rat colon and porcine PYY on a SP-Sephadex ion exchanger were similar. Furthermore, serial dilutions of extracts from the rat colon and porcine PYY had parallel displacement curves in radioimmunoassay. Close intraarterial administration of PYY in cats caused an intestinal vasoconstriction and an inhibition of jejunal and colonic motility. Simultaneously there was a rise in systemic arterial blood pressure. These effects of PYY were also observed after pretreatment with adrenergic blocking agents. It is concluded that PYY is a gastrointestinal peptide that is present mainly in endocrine cells of distal intestine and that has effect on both intestinal motility and the cardiovascular system.

Animals↗
PubMed1977

Expression of endogenous murine leukemia viruses during the course of a protracted immunological disorder.

Mice of the low leukemia (BALB/cJ x A/J)F1 hybrid (CAF1) strain express B-and N-tropic infectious murine leukemia virus (MuLV) after the age of 6 mo. Initation of a protracted immunological disorder, the graft-versus-host reaction (GVHR), at 7 wk of age, accelerates the induction of both these mouse-tropic endogenous viruses, and preferentially enhances the replication of B-tropic MuLV. The earlier appearance of B-tropic MuLV in a greater proportion of mice and in higher titer is thought to be casually related to the eventual development of lymphoreticular tumors in the GVHR mice, since previous studies have shown that these same tumors can be reproduced by inoculating syngeneic recipients with serially passaged GVHR extracts containing B-tropic MuLV.

Age Factors↗
PubMed1995

Telomerase activity in small-cell and non-small-cell lung cancers.

BACKGROUND: Telomerase is an enzyme that adds hexameric TTAGGG nucleotide repeats onto the ends of vertebrate chromosomal DNAs (i.e., telomeres) to compensate for losses that occur with each round of DNA replication. Somatic cells do not have telomerase activity and stop dividing when the telomeric ends of at least some chromosomes have been shortened to a critical length. It has been suggested that immortalized cells (including some, but probably not all, cancer cells) continue to proliferate indefinitely because they express telomerase. PURPOSE: To investigate whether expression of telomerase is a prerequisite for the development of naturally occurring human cancers, we assayed the levels of telomerase activity in specimens of human lung tumor and adjacent normal tissue. METHODS: Using a polymerase chain reaction-based assay, we examined telomerase activity in 136 primary lung cancer tissues and 68 adjacent noncancerous tissues obtained by surgical resection. We also studied telomerase activity in four primary and 23 metastatic lesions obtained through biopsy, (two patients) or autopsy (10 patients). Relative telomerase activity levels were estimated by serial dilutions of extracts prepared from the specimens. Telomerase activity was also assayed in extracts of cells present in pleural fluids from three patients with adenocarcinoma of the lung. RESULTS: Among surgically resected samples, telomerase activity was detected in 109 (80.1%) of 136 primary lung cancer tissues and in three (4.4%) of 68 normal adjacent tissues. All 11 surgically resected specimens of primary small-cell lung cancer (from 11 patients) revealed high levels of telomerase activity, whereas the activity ranged from undetectable to high levels in the 125 surgically resected specimens of primary non-small-cell lung cancer tissue (from 125 patients). Generally, high levels of telomerase activity were observed in metastatic lesions and tumors with altered telomere length. A few primary and, surprisingly, some metastatic tumors did not appear to have detectable telomerase activity. Telomerase activity was, however, detected in cells present in all tested pleural fluids obtained (from three patients with adenocarcinoma of the lung). CONCLUSION: The subset of non-small-cell lung cancers that exhibits only low or undetectable levels of telomerase activity may contain primarily mortal cancer cells. Cancers that exhibit high levels of telomerase activity, such as all of the small-cell lung cancers examined in this study, are likely to consist mainly of immortal cells. IMPLICATIONS: Telomerase activity may be useful both as a diagnostic marker to detect the existence of immortal lung cancer cells in clinical materials and as a target for therapeutic intervention.

Base Sequence↗
PubMed2003

Quantitation of blood lymphocyte mitochondrial DNA for the monitoring of antiretroviral drug-induced mitochondrial DNA depletion.

OBJECTIVE: To investigate the impact of antiretroviral treatment on the mitochondrial DNA (mtDNA) content of peripheral blood mononuclear cells (PBMCs) from HIV-1-infected patients. DESIGN: As absolute mtDNA copy numbers widely differ between individuals, we performed a longitudinal analysis where the patient's first historical specimen was obtained as a baseline reference for relative comparison with subsequent samples from that patient. METHODS: mtDNA and nuclear DNA quantitation per cell (beta-globin gene copies) were both measured by real-time polymerase chain reaction analysis of whole DNA extracts of 361 serial live-cryopreserved PBMCs collected in former trials and clinical follow-ups from 60 individuals with established or recently acquired HIV-1 infections before and during administration of various antiviral combination therapies. RESULTS: mtDNA amounts were stable or increasing over years of natural HIV-1 infection in untreated patients (n = 7), consistent with our finding of a lack of differences in mtDNA copy numbers in patients with either a long established or recent lentivirus infection. Our quantitation system revealed significant changes in mtDNA copy number depending on the designated triple, quadruple, or quintuple anti-HIV drug combinations. Zidovudine + zalcitabine + ritonavir and zidovudine + lamivudine + didanosine regularly lead to mtDNA depletion in each of the treated patients, whereas none of 7 patients (and 35 cell specimens) receiving a stavudine + lamivudine + indinavir combination had any significant mtDNA content variations. In 7 patients, mtDNA copy numbers returned to pretreatment levels and/or higher levels without any interruption of the previously mtDNA-depleting antiretroviral drug combination. CONCLUSION: Our assay system allowed the detection of significant changes in the mtDNA content of PBMCs from HIV-1-infected patients taking antiretroviral drugs, as has been reported in the literature with other detection systems. Yet, mtDNA copy numbers regularly diminished during administration of some but not all nucleoside analog-containing combinations. This, plus the occasional finding that depleted mtDNA contents spontaneously increased to baseline levels and/or higher levels during uninterrupted treatment, should raise a note of caution about resorting to the PBMC mtDNA marker for monitoring of antiretroviral drug-related mitochondrial toxicities.

Acquired Immunodeficiency Syndrome↗