Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Serial Extraction”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,261 records · Page 70Linked to original sources
PubMed1995

Effect of increased intra-abdominal pressure on hepatic extraction and clearance of fentanyl in neonatal lambs.

Fentanyl, an opioid metabolized by hepatic mixed function oxidases, is commonly administered as the primary anesthetic for neonates undergoing surgery. Pharmacokinetic studies have suggested that abdominal surgery in neonates decreases fentanyl clearance, contending that this results from increases in intra-abdominal pressure (IAP) decreasing hepatic blood flow. To examine the effects of IAP on hepatic blood flow and fentanyl clearance, we infused fentanyl to eight neonatal lambs, measured regional blood flows by using the radionuclide-labeled microsphere technique and determined hepatic fentanyl extraction and clearance and hepatic oxygen extraction and consumption at three levels of IAP: 0, 12 and 18 mm Hg. Increased IAP did not affect portal or hepatic blood flow or ductus venosus shunt. Fentanyl extraction was 16.5 +/- 3.0% (mean +/- S.E.) at 0 mm Hg of IAP. Increased IAP decreased hepatic extraction of fentanyl, thereby decreasing fentanyl clearance. Increased IAP did not affect hepatic oxygen extraction or consumption. In two additional animals in which serial measurements of hepatic blood flow were obtained, increased IAP (15 mm Hg) transiently decreased hepatic blood flow with recovery to control values at 2 hr. The authors conclude that fentanyl is poorly extracted by neonatal livers, in contrast to its large extraction ratio in adults. The decrease in fentanyl clearance with increased IAP is consistent with pharmacokinetic studies demonstrating decreased clearance in neonates undergoing abdominal surgery. However, the present study suggests that the mechanism of decreased clearance is decreased hepatic function (decreased fentanyl extraction) rather than decreased hepatic blood flow.

Abdomen↗
PubMed2000

Evaluation of automated nucleic acid extraction devices for application in HCV NAT.

BACKGROUND: To further improve the safety of the blood supply, various national blood transfusion organizations presently use or are in the process of implementing routine HCV NAT in minipools. According to the Committee for Proprietary Medicinal Products (CPMP) of the European Union, the HCV NAT detection limit of the assay should be 100 IU per mL (270 geq/mL) for testing initial plasma pools. Paul Ehrlich Institute (PEI) regulations stipulate that 5000 IU per mL (13,500 geq/mL) must be detected to calculate the amount contributed by individual donations composing the minipool. The sensitivity for HCV RNA extraction achieved by three commercially available laboratory kits was compared. STUDY DESIGN AND METHODS: Nucleic acids from 1-in-3 serial dilutions of an HCV RNA run control (Pelispy, CLB) were extracted with three kits (Cobas Amplicor, Roche Diagnostic Systems; BioRobot 9604, Qiagen; and NucliSens Extractor, Organon Teknika). HCV PCR of all extracts was performed using a second-generation Cobas Amplicor HCV test and the Cobas Amplicor analyzer. RESULTS: The manual Cobas Amplicor, the BioRobot 9604, and the NucliSens Extractor setups allow a 95-percent HCV RNA detection limit of 129, 82, and 12 geq per mL, respectively. The maximal pool size for the manual Cobas Amplicor, the BioRobot 9604, and the NucliSens Extractor kits that would still meet the PEI criteria for HCV NAT in minipools was calculated at 104, 164, and 1125 donations, respectively. CONCLUSION: All three HCV NAT kits evaluated meet the criteria set by CPMP and PEI. The highest sensitivity for HCV NAT screening can be achieved with the high-volume NucliSens Extractor method in combination with the Cobas Amplicor HCV v2.0 test on the Cobas Amplicor analyzer.

Blood Donors↗
PubMed1996

Studies of allergen extract stability: the effects of dilution and mixing.

BACKGROUND: However potent the allergy extracts provided by manufacturers, they are subject to deterioration with storage, especially after dilution or mixture with other extracts. OBJECTIVE: This study was performed to assess separately the deterioration during storage in allergen extract potency caused by dilution or by mixture with allergen extracts that have been reported to contain proteases. METHODS: To assess the effect of dilution, three serial 10-fold dilutions of cat, short ragweed, Bermuda grass, and Dermatophagoides farinae extracts were prepared alone or combined with other extracts. They were stored at 4 degrees C for 3 and 12 months. To assess the effect of mixing with other extracts that have been reported to contain proteases, extracts of timothy grass, Bermuda grass, short ragweed, Russian thistle, white oak, box elder, D. farinae, and cat were stored alone or combined with one or more extracts of American cockroach, Alternaria spp., Cladosporium spp., Penicillium spp., and a house dust mite mix for 3 months at 4 degrees C. RESULTS: Bermuda grass, cat, and house dust mite extracts incurred significant loss of potency at all dilutions with storage. Short ragweed was stable at all dilutions. Potency of extracts of timothy grass, Bermuda grass, Russian thistle, white oak, box elder, and cat were all reduced by combination with one or more extracts potentially containing proteases. Only short ragweed and D. farinae, which was in a final concentration of 25% glycerin, were resistant. Alternaria extract was most frequently responsible for loss of potency, followed by cockroach and Cladosporium extracts. Combination with extracts of Penicillium and a house dust mite mix did not reduce the potency of any extract. CONCLUSIONS: Both dilution alone and mixture with extracts reported to contain proteases caused loss of potency of most extracts tested. Ragweed was uniquely resistant under both conditions of storage.

Allergens↗
PubMed1986

Serial immunologic and histopathologic studies in the treatment of necrotizing fasciitis with combined immunodeficiency by a bovine thymic extract (thymostimulin).

A 25-day-old Chinese female baby developed necrotizing fasciitis (NF) with indurated erythematous patches with superficial necrosis over two thirds of the back. Skin cultures of the lesions yielded Streptococcus pyogenes, enterococcus, Pseudomonas aeruginosa, and Candida species. The patient had lymphopenia with low T cells and T cell subsets. The lymphoproliferative response to mitogen and macrophage migration (MIF) production were also poor. Her lymphocytes were unable to produce IgG in vitro. Also, IgG bearing cells were not demonstrable. The initial lymph node biopsy demonstrated an absence of follicular formation and depletion of lymphocytes in both thymic-dependent and thymic-independent areas. Five weeks after bovine thymic extract (Thymostimulin) treatment, a second lymph node biopsy demonstrated germinal centers containing IgG bearing cells. Three weeks later, a third lymph node biopsy showed germinal centers with cuffs of lymphocytes. Differentiation of the cortex and medulla of the node was demonstrated. IgG was also detectable in an in vitro Ig synthesis study. The total T cells, T cell subsets, lymphoproliferative response MIF production also increased gradually after Thymostimulin treatment. This investigation demonstrated the therapeutic effectiveness of Thymostimulin in NF with underlying combined immuno-deficiency, both serious and frequently fetal diseases, by the histologic and immunologic reconstitution of T and B cell function.

Adjuvants, Immunologic↗
PubMed2003

Narthecium ossifragum (L.) huds. causes kidney damage in goats: morphologic and functional effects.

We studied the effects of Narthecium ossifragum on goat kidneys. Twenty-five Norwegian dairy goats, 5 weeks to 4 months of age, were orally dosed with an aqueous extract from N. ossifragum. In experiment 1, we studied microscopic and functional changes in 12 animals that were euthanatized 2, 3, 4, 5, and 6 days after treatment. In experiment 2, we included ultrastructural studies on serial renal biopsies and urine analysis from five extract-treated animals and two controls. In addition, urine samples were collected from four dosed and two control goats. Ultrasonography revealed perirenal and retroperitoneal fluids. Microscopic changes were observed after 6 hours. The findings, most obvious in the inner cortex and the outer medulla, consisted of cytoplasmic vacuolization, interstitial edema, and focal necrosis of tubular epithelial cells. Ultrastructurally, the tubules had loss of microvilli, irregular cytoplasmic vacuolization, mitochondrial swelling with loss of cristae, and irregular but continuous basement membranes even with necrosis. In the glomeruli, there were occasional endothelial damage and shortening and swelling of the foot processes. Peritubular capillaries had breaks in the vessel walls and irregular endothelial cell edema, and the interstitium had marked edema. The functional lesions included elevated serum urea, creatinine, and magnesium concentrations, a slight decrease in serum calcium concentration, elevated urine protein and urine protein-creatinine ratio, and increased activities of urine alkaline phosphatase and gamma glutamyl transferase. Our findings indicate a fast-acting toxic principle inducing damage by both direct toxic and secondary ischemic effects.

Alkaline Phosphatase↗
PubMed1994

Polymyositis in patients infected with human T-cell leukemia virus type I: the role of the virus in the cause of the disease.

To investigate the mechanism of polymyositis in human T-cell leukemia virus type I (HTLV-I) infection, we studied 6 HTLV-I-positive patients, 3 with polymyositis and 3 with adult T-cell leukemia but without clinical signs of muscle disease, by (a) quantitative single or double immunocytochemistry on serial 4-microns-thick muscle biopsy sections using antibodies to lymphocyte subsets, major histocompatibility complex (MHC) antigens, and HTLV-I proteins; (b) polymerase chain reaction using HTLV-I primers in the RNA and DNA extracted from 50 micrograms of muscle tissue or from serial 5-microns-thick fresh-frozen tissue sections; and (c) cocultures of the patients' HTLV-I-positive peripheral blood lymphocytes with their homologous muscles searching for replication of HTLV-I within the myotubes. In the muscle of patients with HTLV-I-associated myopathy, the predominant endomysial cells surrounding healthy muscle fibers were CD8+ cells followed by CD4+ cells and macrophages. MHC-I antigens were ubiquitous in the muscles of all 6 patients, even in those without endomysial inflammation. HTLV-I sequences were amplified from the whole muscle biopsy specimens but the cells harboring viral antigens were rare endomysial macrophages and not muscle fibers. Although HTLV-I sequences were amplified from all the patients' peripheral blood lymphocytes, these cells did not exert myotoxicity or resulted in viral replication in cocultures with their homologous myotubes.(ABSTRACT TRUNCATED AT 250 WORDS)

Antigens, CD↗
PubMed2006

[An in vitro study on the incidence of the second mesiobuccal canal in the mesiobuccal root of the first and second maxillary molars].

OBJECTIVE: To investigate the incidence of the second mesiobuccal canal (MB2) in the mesiobuccal root of the maxillary first and second molars in Chinese population using three techniques, including the clearing technique, spiral CT scanning and serial root sections. METHODS: A total of 216 extracted human first and 334 second maxillary molars were randomly divided into two groups respectively: group A and B. The teeth in group A were cleared. The specimens in group B were subjected to spiral CT scanning, and then the roots were cross-sectioned every 1 mm from the root apex. Under the Dental operating microscope (DOM), the incidence of MB2 were recorded. RESULTS: (1) The incidence of MB2 in the first and the second maxillary molars were 81.48% and 49.70% respectively by clearing, and 77.78% and 47.31% from S-CT scanning, 88.89% and 53.89% respectively from serial root section. The occurrence of MB2 in maxillary first molars was statistically higher than in maxillary second molars (P < 0.05, chi square test). (2) There was no significant difference among the three approaches for detecting the MB2 canal (P > 0.05, chi square test). CONCLUSIONS: Both the maxillary first molars and the second molars have high incidence of MB2.

Dental Pulp Cavity↗
PubMed2000

Evaluation of automated RNA-extraction technology and a qualitative HCV assay for sensitivity and detection of HCV RNA in pool-screening systems.

BACKGROUND: The objective of this study was the evaluation of NAT technology for the detection of HCV RNA in plasma pools according to the recommendations of the Paul Ehrlich Institute (5000 IU/mL/donation) and the Committee for Proprietary Medical Products (100 IU/mL/manufacturing pool). STUDY DESIGN AND METHODS: Serial dilutions of both the EUROHEP standard (3,800 genome equivalents [geq]/mL; HCV genotype 1) and the World Health Organization (WHO) international standard (100,000 IU/mL; HCV genotype 1) were made in S/D plasma (ESPEP plasma, OctaPharma), which was nonreactive in serologic tests. Serial dilutions of plasma (2 mL) were used for extraction of HCV RNA with an automated version of a nucleic acid isolation method (NucliSens Extractor, Organon Teknika). HCV RNA was co-extracted from 2 mL of plasma, together with 84 copies of an in vitro-synthesized single-strand RNA serving as internal extraction control (IC) to monitor the efficiency of extraction and PCR. Amplification and detection of both HCV RNA and IC RNA were performed with an automated PCR system and a qualitative HCV assay (COBAS Amplicor 2.0 HCV, Roche Diagnostics). RESULTS: A cutoff value of 16 geq per mL (10/10 runs [100% hit rate]) was found by using the EUROHEP standard, whereas the WHO international standard had a cutoff value of approximately 12 IU per mL (10/10 runs [100% hit rate]). The IC had a cutoff value of approximately 17.5 copies per mL (6/6 runs [100% hit rate]). Forty-two copies per mL of IC RNA were found in 282 of 284 runs (99% hit rate). The negative controls (ESDEP plasma) were negative in all experiments. Experiments with pool sizes of 12, 24, 48, and 96 using serial dilutions of the WHO international standard revealed a cutoff value of 8 IU per mL (100% hit rate). The EUROHEP standard and the WHO international standard were detected with a 50 percent detection endpoint of 5.2 geq per mL and 1.5 IU per mL, respectively. CONCLUSION: This test system (NucliSens Extractor, and the COBAS Amplicor 2.0 HCV assay) revealed a high sensitivity for HCV RNA; considering the proposed requirements for sensitivity of NAT assays for the detection of HCV RNA in donor plasma, pool sizes of about 400 donors are possible. These endpoint results indicated that 1 IU is equal to about 3.4 geq.

Autoanalysis↗
PubMed1996

Serial passage of tobacco rattle virus under different selection conditions results in deletion of structural and nonstructural genes in RNA 2.

The RNA genome of tobacco rattle virus (TRV) is bipartite. RNA 2 of the nematode-transmissible TRV isolate PPK20 encodes the viral coat protein (cp) and proteins with molecular weights of 29,400 and 32,800 (29.4K and 32.8K proteins). When this isolate was serially passaged in tobacco by using phenol-extracted RNA as the inoculum in each transfer, defective interfering (DI) RNAs rapidly accumulated. A number of these DI RNAs were cloned. Six DI RNAs had single internal deletions in RNA 2 that removed most of the cp gene, the 29.4K gene, and the 5' half of the 32.8K gene. The borders of the deletions in these DI RNAs were found to be flanked in the genomic RNA 2 by short nucleotide repeats or sequences resembling the 5' end of TRV genomic and subgenomic RNAs. Two DI RNAs were found to be recombinants containing a 5' sequence derived from RNA 2 and a 3' sequence derived from RNA 1. When serial passage of TRV isolate PPK20 was carried out by using leaf homogenates as inocula in each transfer, accumulation of a DI RNA (designated D7) with a functional cp gene was observed. The deletion in D7 covered the 3' end of the cp gene, the 29.4K gene, and the 5' half of the 32.8K gene. An infectious cDNA clone of D7 RNA was made. In mixed infections, D7 RNA rapidly outcompeted RNA 2 but did not compete with RNA 1. The deletion in D7 RNA abolished the nematode transmissibility of the PPK20 isolate. These results may explain the observation that many laboratory isolates of tobraviruses have lost their nematode transmissibility and contain RNA 2 molecules of widely different lengths.

Culture Media↗
PubMed2006

A study of periapical lesions correlating the presence of a radiopaque lamina with histological findings.

OBJECTIVE: To compare the presence or absence of a radiopaque lamina of 60 human periapical lesions with the histological findings from a case study in dental practice. STUDY DESIGN: Paralleling radiographs were taken of 60 teeth with periapical radiolucencies. The periapical radiographs were scanned, and standardized images were evaluated on a computer screen for the presence or absence of a radiopaque lamina by 2 calibrated observers according to agreed criteria. The serially sectioned histological specimens were obtained from the extraction of 13 endodontically treated teeth and 27 nontreated teeth, as well as 20 post-treatment teeth with apical periodontitis removed during periapical surgical procedure; the specimens were classified according to agreed criteria. The findings of the 2 evaluations were compared. RESULTS: Out of 57 electronic images that could be interpreted, 10 lesions had a radiopaque lamina, but of these only 3 were histologically diagnosed as cysts, while 7 were granulomas or abscesses. Out of 47 lesions without a radiopaque lamina, 40 were histologically diagnosed as granulomas or abscesses, while 7 were cysts. CONCLUSIONS: The diagnosis of periapical lesions cannot be made on the basis of the presence or absence of a radiopaque lamina, but requires histological examination of serial sections.

Diagnosis, Differential↗
PubMed1994

Testing garlic for possible anti-ageing effects on long-term growth characteristics, morphology and macromolecular synthesis of human fibroblasts in culture.

The beneficial effects claimed for the use of garlic as a nutritional supplement include detoxification, antioxidation, antifungal activity, antibacterial activity, tumour suppression and, possibly, anti-ageing and rejuvenating effects. We have used the Hayflick system of cellular ageing in culture in order to test garlic for its anti-ageing effects on long-term growth characteristics, morphology and macromolecular synthesis of human skin fibroblasts. Our results show that an addition of garlic extract into the normal cell culture medium can support serial subculturing for over more than 55 population doublings in 475 days, and that this treatment has some youth-preserving, anti-ageing and beneficial effects on human fibroblasts in terms of maximum proliferative capacity and morphological characteristics. In comparison, similar or lesser doses of garlic extracts are growth inhibitory for cancerous cells that could not be grown over longer periods in the presence of garlic. To our knowledge, this is the first report of the effects of garlic on the long-term growth characteristics and macromolecular synthesis of normal human skin cells, the results of which have applications for both anti-ageing and anti-cancer research.

Adult↗
PubMed1988

Biotinylated and radioactive DNA probes for detection of varicella-zoster virus genome in infected human cells.

We have developed and compared two DNA dot hybridization methodologies with similar probes (radioisotope-labelled and biotin-labelled) to detect varicella-zoster virus (VZV) DNA in three different cell cultures at varying times post-infection. Control cultures included uninfected monolayers of the same cells. Cellular DNA was isolated by a standard phenol extraction method, after which the DNA was quantified, serially diluted and blotted onto nitrocellulose or nylon membranes. The VZV DNA probe, which consisted of the large Hind III A fragment (27 of the total 125 kbp), was produced in two separate nick translation systems. The first contained 100 microCi [32P] and 0.4 microgram Hind III fragment A of the varicella genome, while the second probe employed a biotin-7-dATP analogue and 1.0 microgram of the Hind III fragment A. Direct visualization on the membrane or the exposed radiographic film showed a dot of varying intensity whenever viral genome was detected with either the biotin or the radioactive probe, respectively. With the [32P]-labelled probe, we detected VZV genomic sequences within 0.5 microgram total DNA at 12 h post-infection. This amount corresponded to approximately 5-10 pg of viral DNA. By comparison, hybridization with the biotin-labelled probe required 0.5-1.0 micrograms total DNA from infected cells. Similar tests on DNA extracted from uninfected cell samples were negative with both probes.

Biotin↗
PubMed2003

The use of extraction chromatography resins to concentrate actinides and strontium from soil for radiochromatographic analyses.

An analytical technique utilizing selective extractant resins to concentrate strontium and actinides from soil followed by separation with radiochromatography was evaluated. The technique was tested using uncontaminated soil samples spiked with a radionuclide tracer solution that were either microwave-aided acid digested or leached with a strong acid. Extraction of the strontium and actinides from the acidified solution was accomplished using a serial arrangement of Sr-Resin and TRU-Resin columns. The combined eluate solutions from the extraction resins were treated with HNO(3) and H(2)O(2) to oxidize residual extractant and eluates prior to separation and analysis of the radionuclides by radiochromatography. Chromatograms obtained with larger soil mass loadings resulted in either incomplete peak resolution of the tracers or had highly variable peak elution times, indicative of an ionic interfering constituent(s). Better separations (e.g., chromatograms that resolved all radioactive constituents) were obtained when the sample mass loading was decreased, but with a concurrent decreased sensitivity for the radionuclides. Elemental analyses of the soil were conducted to provide data on the ionic constituents in unprocessed soil and post-processed soil samples. These results identified aluminum as an interfering contributor to the poor performance exhibited by the radiochromatographic separations.

Actinoid Series Elements↗
PubMed1984

The pharmacology of diaziquone given in intravenous or intracarotid infusion to normal and intracranial tumor-bearing puppies.

Diaziquone (also called "aziridinyl benzoquinone," or AZQ), an antitumor drug designed to penetrate the blood-brain barrier, has demonstrated activity against central nervous system (CNS) neoplasms. Four-hour infusions of carbon-14 (14C)-labeled AZQ (0.8 mg/kg) were given via the left common carotid artery or left brachial vein to two groups of puppies. A third group, harboring a transplantable canine glioma, received 14C-AZQ by intravenous infusion. Levels of chloroform (CHCl3)-extractable 14C (AZQ only) and total 14C (AZQ and metabolites) were determined in serial samples of plasma and cerebrospinal fluid (CSF). At the end of the infusion time, total and CHCl3-extractable 14C levels were determined in brain and tumor. Intra-arterial infusion of AZQ caused no histological abnormalities in the retina or brain. For the intravenous infusion group, the concentrations of CHCl3-extractable 14C (in nmol/ml or nmol/gm) were 0.68, 0.35, and 0.84 for plasma, brain, and CSF, respectively. For the intra-arterial infusion group, the concentrations were 0.25, 0.13, and 0.32 for plasma, brain, and CSF, respectively. Comparison of right and left hemispheres following intra-arterial infusion showed a slightly higher concentration of 14C in the ipsilateral (left) hemisphere, with concentrations (nmol/gm) of CHCl3-extractable 14C/total of 14C of 0.15/0.87 on the left and 0.12/0.65 on the right. Concentrations (nmol/gm) of CHCl3-extractable 14C/total 14C in brain and tumor were 0.60/1.24 and 0.58/1.65, respectively. In tumor-bearing animals, tumor and surrounding brain contained similar concentrations of AZQ, but there were higher concentrations of metabolites in tumor. This may reflect different metabolism of AZQ within brain and tumor or different permeability to metabolites. This study revealed that AZQ enters the CNS and brain-tumor tissue in substantial concentrations and that there is no significant advantage to intracarotid infusion of AZQ.

Animals↗
PubMed2003

Downregulation of eNOS mRNA expression by TNFalpha: identification and functional characterization of RNA-protein interactions in the 3'UTR.

OBJECTIVE: We have previously shown that downregulation of endothelial nitric oxide synthase (eNOS) expression by tumour necrosis factor-alpha (TNF alpha) resulted entirely from the marked destabilization of the eNOS mRNA. As the 3'-untranslated region (3'UTR) in many eukaryotic mRNA has been well documented to bind regulatory trans-factors in the control of transcript stability, we have examined protein binding to this region of the eNOS mRNA. A high degree of homology amongst human and bovine 3'UTR also suggests that important functional features that are conserved through evolution are present within this region. METHODS: RNA-protein interactions were studied in cross-linking assays, in which radiolabelled RNA encoding the human eNOS 3'UTR or selected sequences was incubated with cytoplasmic extracts of cultured human umbilical vein endothelial cells (HUVECs). Serial 5'- and 3'-truncated deletional mutations of the eNOS 3'UTR were generated to identify the specific binding sequences. eNOS mRNA expression in HUVECs was assessed by RT-PCR analysis. RESULTS: Using radiolabelled RNA encoding the entire 418-nucleotide 3'UTR, we have identified ribonucleoprotein complexes (RNPs) of approximate molecular weights of 53, 56 and 66 kDa in the endothelial extracts. The formation of the 53- and 56-kDa RNPs was upregulated by TNF alpha, while the formation of the 66-kDa RNP was downregulated. Formation of the 53-kDa RNP was favoured by RNA fragments that contained sequences from the proximal and distal portions of the 3'UTR, whereas the formation of the 66-kDa RNP was favoured by RNA fragments with the AU-rich distal end. RNA fragments containing a CU-rich 158-nucleotide sequence from the medial portion of the eNOS 3'UTR (designated M158) favoured the formation of the 56-kDa RNP. Adenoviral gene transfer and overexpression of M158 RNA, as a protein-binding decoy to prevent the formation of the 56-kDa RNP on the endogenous transcripts, attenuated the TNF alpha-induced downregulation of eNOS mRNA in cultured endothelial cells. CONCLUSION: Our results demonstrate that the regulation of eNOS expression involves the specific binding of cytoplasmic proteins to highly conserved elements along the 3'UTR, and the 56-kDa RNP represents a novel regulatory trans-factor in the destabilization of eNOS transcripts.

3' Untranslated Regions↗
PubMed1992

Serial cultivation of normal human keratinocytes: a defined system for studying the regulation of growth and differentiation.

We have developed a defined method for human epidermal keratinocyte culture. The minimally supplemented basal medium supported establishment of primary cultures from neonatal foreskin in a defined environment. It also supported serial cultivation and rapid expansion of cell number. Casein replaced serum for defined cryopreservation. Cells were serially cultivated in medium containing 0.08 mM calcium. The rate of cell division however remained high after addition of 1.8 mM calcium. The particulate transglutaminase activity of the cultures was low at confluence, even in the presence of 1.88 mM calcium, indicating an enrichment of the basal cell population. Culture with small amounts (0.3%) of chelated serum increased particulate transglutaminase activity approximately 2.2-fold in low calcium cultures and approximately 3.5-fold in high calcium cultures. A gradual reduction in growth rate of serum-treated cultures upon serial cultivation also indicated a depletion of cells with basal cell character. Bovine hypothalamic extract and cholera toxin were able to avert, in part, the differentiation-promoting effects of serum. Keratinocytes serially cultivated in the defined medium maintained the ability to develop normally into a morphologically differentiated epidermis.

Blood Proteins↗
PubMed2004

Transcriptome of mouse uterus by serial analysis of gene expression (SAGE): comparison with skeletal muscle.

The aim of this study was to identify the transcriptome of the normal mouse uterus by Serial Analysis of Gene Expression method. mRNA was extracted from the uterus and also from the gastrocnemius muscle of mice. Short sequences (tags), each one usually corresponding to a distinct transcript, were isolated and concatemerized into long DNA molecules which were cloned and sequenced. We detected 44,484 tags for the uterus and 42,518 tags for the muscle, representing 14,543 and 14,958 potential transcript species, respectively. Seventy-five and sixty-nine genes were expressed at more than 0.1%, thus corresponding to 37 and 34% of the mRNA population detected in the respective tissues. In both cases, the most highly expressed genes are especially involved in muscle contraction, energy metabolism, and protein synthesis. Compared to skeletal muscle, some differentially expressed genes in the uterus are likely to correspond to its specific reproductive functions. The majority of these genes remain to be characterized. More than 70% of the different tags detected in the uterus did not match any sequence in the public databases and can represent novel or poorly identified genes. This study is the first quantitative description of the transcriptome of the uterus.

Animals↗