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At least 127 records · Page 7Linked to original sources

PubMed1999

Expression profiling by iAFLP: A PCR-based method for genome-wide gene expression profiling.

The availability of comprehensive sets of genes has prompted the researchers to carry out systematic collection of gene expression data. RT-PCR has the highest specificity and sensitivity for transcript detection among all available methods. Low throughput, especially when quantitative data are desired, has precluded RT-PCR from genome-wide application. Here we report a PCR-based expression profiling method, introduced amplified fragment length polymorphism (iAFLP), that has the same specificity and sensitivity as RT-PCR and a throughput level comparable to that of DNA-microarray hybridization. In this method, restricted ends of total cDNAs from six sources were ligated with adaptors having various length of short insertions to a common sequence (polymorphic adaptors). Amplification of a pool of these differentially adapted cDNAs with a gene-specific primer and an adaptor primer allows us to quantitate the abundance of any transcript in six mRNA sources. Using three different primer colors this technique allows quantitation of expression of 864 genes across six different sources per day with a single autosequencer, which is comparable to the throughput of microarray analysis in terms of number of genes x number of sources.

Animals↗

PubMed2006

Genetic profiling of dendritic cells exposed to live- or ultraviolet-irradiated Chlamydia muridarum reveals marked differences in CXC chemokine profiles.

Chlamydia trachomatis is a major cause of sexually transmitted disease worldwide for which an effective vaccine is being actively pursued. Current vaccine efforts will be aided by elucidating the interaction between Chlamydia and dendritic cells (DCs). Protective immunity appears to develop slowly following natural infection in humans, and early vaccine trials using inactivated C. trachomatis resulted in partial, short-lived protection with possible enhanced inflammatory pathology during re-infection. Thus, immunity following natural infection with live chlamydia may differ fundamentally from immune responses induced by immunization with inactivated chlamydia. We explored this conjecture by studying the response of DCs exposed to either viable or inactivated [ultraviolet (UV) -irradiated] chlamydia elementary bodies (EBs; designated as Live-EB and UV-EB, respectively) using Affymetrix GeneChip microarrays. Thirty-one immunologically characterized genes were differentially expressed by DCs following exposure to Live-EB or UV-EB, including two glutamic acid-leucine-arginine cysteine-X-cysteine (ELR CXC) neutrophil chemoattractant chemokines, Cxcl1 (KC), and Cxcl2 (MIP-2). Up-regulation of these genes by Live-EB as compared to UV-EB was verified by quantitative reverse transcription-polymerase chain reaction and increased chemokine secretion was confirmed by enzyme-linked immunosorbent assay both in vitro and in vivo. Immunofluorescence and fluorescence-activated cell sorter analysis of chlamydia-infected lung tissue confirmed that Live-EB but not UV-EB induced significant DC and neutrophil infiltration during infection. These observations demonstrate that the development of an antichlamydial immune response is dramatically influenced by chlamydial viability. This has implications as to why early inactivated chlamydial vaccines were ineffective and suggests that new vaccine design efforts may benefit from in vitro DC screening for ELR chemokine expression profiles.

Animals↗

PubMed2006

Effects of antidepressant treatment on gene expression profile in mouse brain: cell type-specific transcription profiling using laser microdissection and microarray analysis.

A gene expression study of mice treated with the tricyclic antidepressant amitriptyline was performed. To enable the detection of cell type-specific expression changes, laser-microdissected nucleus accumbens was analysed after 4 and 28 days of treatment. After 4 days of treatment no significantly regulated genes could be detected in this study. In contrast, 95 genes exhibited different expression levels in animals treated for 28 days with amitrityline compared with sham animals. This observation reflects the long-term effects and adaptation processes observed in patients treated with this drug. Among the regulated genes are receptors belonging to the dopamine-dependent signalling cascade, ion channels (mainly voltage-dependent potassium and calcium channels) potentially involved in signalling cascades and neuropeptides. The results support the hypothesis that the therapeutic effect of this antidepressant is much more complex and not confined to a reuptake inhibition of neurotransmitters. Paradigms inducing only weak expression changes, which may be limited to certain cell types within the highly complex brain structure, can therefore be reliably investigated by applying a cell type-specific expression profiling technique based on laser microdissection and subsequent RNA amplification followed by DNA microarray analysis.

Amitriptyline↗

PubMed1982

Functional ability profiles for driver licensing. Exemplification by visual profile.

The impaired driver is evaluated for driver licensing according to a functional ability profile. This test correlates, in increments, the physical or medical ability of the driver with the risk categories and the vehicles the driver is permitted to operate. The person is responsible for his fitness to drive. The physician may help in patient evaluation by participation in a medical advisory board or in public education. Guidelines for 11 areas of potential driving impairments were developed in the state of Utah. These principles are illustrated with visual standards.

Automobile Driver Examination↗

PubMed1983

Acute effects of steroids on immune complex profile of patients with systemic lupus erythematosus. Correlation of profile with development of target organ involvement.

Eleven patients with active systemic lupus erythematosus, previously untreated, were studied to 1) determine the acute effect of corticosteroids on circulating immune complex (CIC) levels and 2) correlate the initial CIC profile with the development of organ system involvement. Using serial measurements of CIC as detected by assays for cryoglobulins and binding to C1q, Raji cells, and rheumatoid factor, we found that levels of CIC change little during the first month of high dose daily steroid therapy, but they uniformly decrease to near normal by 6 to 12 months. High levels of CIC detected by Raji cell assay early in the course of systemic lupus erythematosus and before steroid therapy appear to be predictive of the development of chronic lupus nephritis (P less than 0.005).

Adult↗

PubMed2002

Functionally active estrogen receptor isoform profiles in the breast tumors of African American women are different from the profiles in breast tumors of Caucasian women.

BACKGROUND: Several cancer surveys have shown that African-American women (AAW) develop highly aggressive breast tumors and experience about three times higher mortality rates compared with other populations. Generally, breast tumors in AAW are poorly differentiated or undifferentiated and exhibit increased frequency of nuclear atypia, higher mitotic activity, higher S-phase fraction, and tumor necrosis. The molecular factors responsible for these tumor characteristics are mostly unknown. METHODS: To explore whether the aggressive tumor biology observed in AAW is related to distinct alterations in estrogen receptor (ER) isoforms, the relative expression levels of four functionally active ER isoform mRNAs, ERalpha wild type, ERbeta wild type, ERalpha exon 3delta, and ERalpha exon 5delta, were measured by reverse transcriptase-polymerase chain reaction analysis in 18 immunohistochemically ERalpha positive tumors and in 6 ERalpha negative tumors and their matched normal tissues. RESULTS: In the tumors of AAW, the protective ERbeta isoform was decreased significantly compared with matched normal tissues (paired t test; n = 24 patients; P = 0.0018). In addition, both the constitutively active ERalpha exon 5delta and the dominant negative ERalpha exon 3delta mRNA levels were elevated in tumor tissues compared with matched normal tissues (paired t tests; n = 24 patients; P = 0.0002 and P = 0.024, respectively). CONCLUSIONS: The data presented here show for the first time that functionally active ER isoform profiles in the breast tumors of AAW are different from those in Caucasian women. The tumors in AAW are characterized by decreased levels of the protective ERbeta isoform and elevated levels of the constitutively active ERalpha exon 5delta isoform. Variations in estrogen-mediated signaling because of the alterations in these two ER isoforms may account in part for differences in tumor biology between AAW and Caucasian women.

Black People↗

PubMed2003

Transmission profile of hepatitis B virus infection in the Batam region, Indonesia. Evidence for a predominantly horizontal transmission profile.

The aim of this study was to describe the distribution of hepatitis B virus markers among the autochthonous and immigrant multicultural populations of Riau province, Indonesia, in order to define the groups at risk and their infrastructure. This investigation was part of a large hepatitis B vaccination study. A total of 9701 healthy individuals, aged 5 years or older and living in the urbanised area of Batam, near Singapore, and on the surrounding islands were included. Socio-epidemiological data were collected, blood was drawn, and sera were tested for antibodies to hepatitis B core (anti-HBc) and surface antigen (anti-HBs). Anti-HBc-positive sera were tested against hepatitis B surface antigen (HBsAg). All tests comprised immunoassays from Roche (Germany) using Elecsys 2010. Complete data were available from 9314 subjects. The results showed relatively low prevalences of anti-HBc (a marker of a previous hepatitis B infection) and HBsAg in the 5-year-old chiLdren (5.8 and 1.9%, respectively) that increased continuously with increasing age. High anti-HBs levels (>1000 IU/L) were found in all age cohorts, indicating a lifelong threat of active hepatitis B infection. In conclusion, the transmission profile of hepatitis B appeared to be mainly horizontal (person-to-person) in the highly endemic region studied. Vertical transmission was less than 5%. The horizontal transmission routes included non-sexual activities of life since children <10 years of age also showed considerable infection rates. The results underline the need for catch-up hepatitis B vaccination programs for children and adults.

Adolescent↗

PubMed1996

The Diabetes Health Profile (DHP): a new instrument for assessing the psychosocial profile of insulin requiring patients--development and psychometric evaluation.

The aim of the studies was to evaluate the psychometric properties and construct validity of the Diabetes Health Profile (DHP-1). Content for the DHP-1 was derived following in-depth interviews with 25 insulin dependent and insulin requiring patients, a review of the literature and discussions with health care professionals. Initial analysis of the factor structure of the DHP-1 was carried out on the responses of 239 insulin dependent and insulin requiring patients, with a mean age of 40.85 years (SD = 13.0), resulting in a 43 item three factor solution. The 43 item version of the DHP-1 was completed by 2,239 insulin dependent/requiring patients (mean age = 39.8, SD = 10) years. Fifty-one per cent were men. A forced three factor Principal Factoring Analysis with varimax rotation was carried out. Eleven items were excluded with item factor cross loadings > 0.30 or item factor loadings < 0.30. PAF analysis of the 32 items resulted in a three factor solution accounting for 33% of the total explained variance. The three factors were interpreted as Psychological Distress, Barriers to Activity and Disinhibited Eating. Factor congruence between subsamples were: Psychological distress (0.93), Barriers to Activity (0.93) and Disinhibited Eating (0.99). Coefficients of congruence between men and women were 0.94, 0.92 and 0.99 for Psychological Distress, Barriers to Activity and Disinhibited Eating respectively. Internal consistency of the three factors (Cronbach's alpha) were: Psychological Distress (0.86), Barriers to Activity (0.82), and Disinhibited Eating (0.77). Construct-convergent validity was investigated on a sample of 233 insulin dependent and insulin requiring patients (mean age = 51.46 years). Psychological Distress and Barriers to Activity subscales correlated with the Hospital Depression and Anxiety Scale = 0.50 to 0.62, p < 0.01) and subscales of the SF-36 (range: r = -0.17 to -0.62, p < 0.01). These findings lend support to the construct validity and reliability of the DHP-1 and that it is suitable for further development.

Adult↗

PubMed1977

A comparison of the differential DNA melting profiles with the CsCl density profiles of DNA from Escherichia coli, cow, mouse, rat and chicken.

Moderate resolution thermal denaturation profiles are presented for the purified DNAs from Escherichia coli, cow, mouse, rat and chicken. All show multiple thermal transitions indicative of large blocks of DNA with very similar base composition. The eucaryotes all have much more of this kind of DNA than does E. coli. The satellite DNAs of cow and mouse are clearly visible and it is likely that the other transitions represent additional families of repeated DNA.

Animals↗

PubMed1993

Analysis of a Danish Caucasian population sample of single locus DNA-profiles. Allele frequencies, frequencies of DNA-profiles and heterozygosity.

The frequency distributions of the length of restriction fragments (HinfI) revealed by RFLP-analysis (restriction fragment length polymorphism) of blood samples from 482 Danish Caucasians using the single locus VNTR (variable number of tandem repeats) probes MS1, MS31, MS43a and YNH24 are reported. From two blood samples three fragments were obtained with MS1. The consistency of the characteristic allele frequency distribution for each probe is exemplified by comparing the accumulated frequency curves obtained with MS43a in samples consisting of 50 and 920 bands, respectively. The distribution of the differences in migration distance for the two fragments of a bandpair was investigated. The results suggest that the high frequency of apparent homozygotes observed is due mainly to coalescence of close heterozygotes. The distribution of frequencies of 437 DNA-profiles is reported.

Bias↗

PubMed1991

Estimation of impurity profiles of drugs and related materials. Part VIII: Combined application of high-performance liquid chromatography and NMR spectroscopy in the impurity profiling of drugs.

The usefulness of the joint application of HPLC and NMR spectroscopy in drug impurity profiling is demonstrated by the following examples: (1) identification of Z and E isomers of 17 alpha-ethynyl-4-oestrene-3 beta, 17-diol-3-acetate-17-(3'-acetoxy-2'-butenoate) in ethynodiol diacetate; (2) identification of the p-tolyl analogue as the impurity of enalapril maleate; (3) identification and quantification of 2'-dehydro-pipecuronium bromide in pipecuronium bromide. The possibilities of utilizing NMR spectroscopy for the identification and quantification of the impurities with and without their isolation are discussed.

Androstane-3,17-diol↗

PubMed2005

Focused metabolomic profiling in the drug development process: advances from lipid profiling.

The highly parallel analytical technologies comprising 'omics promised to dramatically improve drug development efficiency by increasing knowledge and improving decision-making capabilities. On this point, the 'omics have largely been a disappointment. The major reason genomics, transcriptomics and proteomics fail to improve decision making capabilities is that they produce so many false positive results that it is difficult to be sure that findings are valid. Metabolomics is not immune to this problem but, when practiced effectively, the technology can reliably produce knowledge to aid in decision making. In particular, focused metabolomics platforms - those that restrict their target analytes to those measured well by the technology - can produce data with properties that maximize sensitivity and minimize the false discovery problem. The most developed focused metabolomics area is lipid profiling.

Animals↗

PubMed2001

Unique phenotypic profile of monocytoid B cells: differences in comparison with the phenotypic profile observed in marginal zone B cells and so-called monocytoid B cell lymphoma.

Monocytoid B cells (MBCs) are a subset of B cells that may be recognized in several reactive and tumoral lymph node conditions, including toxoplasmic lymphadenitis, infectious mononucleosis, and Hodgkin's lymphoma. Although this is a commonly observed cell population, which has even given its name to a type of lymphoma, MBC lymphoma, scarcely any information is available about the function and characteristics of this cell type. A relationship with marginal zone (MZ) B lymphocytes has been claimed for MBCs, but this has not yet been fully proven. Indeed, specific markers for MBCs are still lacking, which has made it difficult to analyze their relationship with other B cell subpopulations and confirm the existence of tumors deriving from this B cell subset. We used a panel of cell cycle markers to explore the characteristics of MBCs and their relationship with MZ B cells, nodal MZ lymphoma, and splenic MZ lymphoma. We therefore compared the phenotypic profile of MBCs in different conditions with normal MZ B cells within the spleen and mesenteric lymph nodes, with a group of seven cases of nodal MZ/MBC lymphoma and another group of five cases of splenic MZ lymphoma. MBCs were mainly in the G(0) to G(1) phases, as deduced from the presence of a proportion of between 10 and 35% Ki67-positive cells, whereas very low expression was observed with cyclin A and cyclin B staining. Nests of MBCs were clearly labeled by the expression of p21(WAF1), a cyclin-dependent kinase inhibitor (CKI), rarely detectable in benign lymphocytes, and by cyclin E. Basically all MBCs were bcl-2-negative, and high cyclin D2 and cyclin D3 were also detected in these cells, at proportions and intensities above expected levels, when the percentage of proliferating cells was taken into account. p27(KIP1) expression was characterized by homogeneous reactivity, higher than that observed in other B cell populations with a relatively high-growth fraction. Immunoglobulin staining showed undetectable light and heavy chains. However, splenic MZ cells, nodal MZ lymphoma, and splenic MZ lymphoma showed a distinct expression of IgM and bcl-2, with high p27 (KIP1) nuclear expression and undetectable or low levels of cyclin A, B, E, or D, or p21(WAF1) expression. The data from this study show an unexpected immunophenotype in MBCs, different from the one observed in splenic and lymph node MZ B cells. This suggests that either MBCs are a unique B cell population from a distinct cell lineage, or if related to MZ cells, they would represent a definite differentiation stage characterized by a distinctive immunophenotype. They also show so-called MZ/MBC lymphoma to be more closely related to lymph node and splenic MZ B cells, as they do not share the most distinctive features of MBCs.

B-Lymphocytes↗

PubMed1980

Studies of female urethral pressure profile. Part I. The normal urethral pressure profile.

The urethral pressure profile is influenced by the degree of patient relaxation, which explains marked variations in measured pressure values among normal subjects, as well as the frequent lack of reproducibility in a given subject. The quality of relaxation can be estimated by observing the difference between maximum resting and maximum holding pressures. The most reproducible parameters are 1) the total urethral length, 2) the length to the point of maximum pressure and 3) the ratio of area to the point of maximum pressure over total area. Changes in intravesical volume do not produce significant differences in peak pressure, total area or area to maximum pressure. When subjects were asked to strain the pressure in the proximal urethra increased by 60% of the intravesical pressure observed, even in the absence of a vesical neck opening.

Adult↗

PubMed2002

Estimation of impurity profiles of drugs and related materials: part XXI. HPLC/UV/MS study of the impurity profile of ethynodiol diacetate.

The impurity profile of ethynodiol diacetate was investigated using the HPLC/UV/MS method. Using the slightly modified HPLC method of USP 24 two impurities, earlier isolated by preparative HPLC and investigated by NMR spectroscopy were separated and characterised. The mass spectra amended by the diode-array UV spectra supported the earlier found structures (E and Z isomers of 17alpha-ethinyl-estr-4-ene-3beta,17-diol-3-acetate-17-(3'-acetoxy-2'-butenoate). Another, hitherto not described impurity, 17alpha-ethinyl-estr-4-ene-3beta,17-diol-3-acetate-17-(3-oxo-butanoate) has also been separated and characterised by means of its mass spectrum, NMR and UV spectra.

Chromatography, High Pressure Liquid↗

PubMed1995

Correlation of change in phage type with pulsed field profile and 16S rrn profile in Salmonella enteritidis phage types 4, 7 and 9a.

Using pulsed-field gel electrophoresis (PFGE) and 16S rRNA (rrn) analysis (ribotyping), the in vivo derivation of strains of Salmonella enteritidis PTs 9a and 7 from a strain of S. enteritidis PT 4 has been demonstrated. All strains were isolated from a single patient over a 6-week period. Further studies have demonstrated that in terms of pulsed-field profile and ribotype, the genotypes of the patient-derived strains differed from those of the reference strains of the respective phage types. It is concluded that when used in combination, these methods can provide evidence of phylogenetic relationships in apparently unrelated S. enteritidis phage types isolated during pathogenesis of disease.

Bacteriophage Typing↗

PubMed1975

[The relationship between age and the cerebrospinal fluid protein profile profile of normal children. Cellulose acetate and agarose gel electrophoretic studies].

Neuropädiatrie 6: 383--397 (1975). CSF samples of 63 normal children 2 days to 14 years of age were concentrated (ultrafiltration in "Kollodiumhülsen") and examined by electrophoresis on cellulose acetate and agarose gel. In the CSF of all children 7 protein fractions were found by cellulose acetate electrophoresis. Analysing the pherograms by the means of an analog computer 15 fractions could be separated by agarose gel electrophoresis. Compared with the children of school age the newborn showed low prealbumin, beta1-globulin and tau-globulin fractions besides high albumin and gamma-globulin concentrations owing to an increased permeability of the blood-CSF barrier. During the following weeks a low prealbumin and a high albumin fraction still was found. Children from 3 month to 2 years of age showed a moderate increase of the relative prealbumin and beta1-globulin concentration in comparison with older children. The gamma-globulin fraction was lowest beyond the third month of life and increased again in the group of the 2--14 years old children. The variation of the protein profile with age could not only be demonstrated by the more difficult agarose gel electrophoresis but also by a simple technique of cellulose acetate electrophoresis.

Adolescent↗

PubMed1991

Urine profiles and kidney histology following intravenous diatrizoate and iohexol in the degeneration phase of gentamicin nephropathy in rats. Effects on urine and serum profiles.

Urine chemical profiles were followed for three or nine days after intravenous injection of diatrizoate, iohexol, or saline in 30 rats, where a tubulointerstitial nephropathy was induced by gentamicin given over an eight-day period. Another ten rats injected with saline served as controls. Compared to injection of saline, both iohexol and diatrizoate induced dysfunction. The excretion of the cytoplasmic enzyme lactate dehydrogenase was significantly greater following iohexol than following diatrizoate. No significant differences between the two media were shown by the various serum components examined. Among the gentamicin-treated rats, light microscopy showed prolonged occurrence of tubular necrosis and a more intensive round cell infiltration following iohexol than following diatrizoate and saline. Both contrast media induced further temporary renal dysfunction in rats with gentamicin nephropathy; iohexol induced more morphologic changes than diatrizoate.

Animals↗