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PubMed2001

Studies on characterisation and thermal behaviour of 3-amino-5-nitro-1,2,4-triazole and its derivatives.

3-Amino-5-nitro-1,2,4-triazole (ANTA) and its derivatives have been prepared in order to carry out the systematic studies on structural aspects, explosive and thermal behaviour. Thermal studies were carried out for ANTA and 4,6-bis-(3-amino-5-nitro-1H-1,2,4-triazole-1-yl)-5-nitropyrimidine (DANTNP) by thermogravimetry (TG), differential thermal analysis (DTA), differential scanning calorimeter (DSC) and manometric thermal analysis. The results show that (DANTNP) is more thermally stable than ANTA when compared in terms of activation energy.

Explosions↗
PubMedDate not supplied

Enterocyte response to ischemia is dependent on differentiation state.

Enterocytes at the tips of microvilli are more sensitive to an ischemic insult than those cells residing in the crypts, an effect thought to be due to a relative lack of collateral flow. We speculated that this increased cellular sensitivity to ischemia might be an intrinsic feature of the cells related to their differentiated phenotype. To test this hypothesis, enterocyte response to ischemia was determined using both in vivo and in vitro models. For the in vivo studies, male Sprague-Dawley rats underwent laparotomy, and small intestinal ischemia was induced by clamping the superior mesenteric artery for 30 or 60 minutes, after which reperfusion was allowed for various time points up to 4 days. Injury was assessed histologically, as well as with Northern blots, probing for the enterocyte differentiation markers intestinal alkaline phosphatase and lactase, as well as the gut-epithelial marker villin. Mucosal changes consistent with ischemia/reperfusion injury were evident--that is, a rapid inflammatory response followed by progressive villus cell loss beginning at the tips and progressing to the crypts, depending on the degree of insult, with an eventual return to normal microanatomy. Intestinal alkaline phosphatase and lactase were lost immediately after ischemia and returned with reperfusion, confirming that the differentiated cells are particularly sensitive to ischemic injury. The in vitro studies employed two separate models of enterocyte differentiation: sodium butyrate-treated HT-29 cells and Caco-2 cells maintained for 7 days after confluence. In both models, undifferentiated and differentiated cells were subjected to treatment with 2-deoxyglucose and oligomycin-A (in vitro model of ischemia) and apoptosis was assessed by fluorescence-activated cell sorting analysis. Differentiation of both cell lines resulted in a significantly greater apoptotic response to ischemia compared to undifferentiated cells exposed to an identical insult. We conclude that differentiated enterocytes may be inherently more sensitive to ischemia-induced injury than their undifferentiated counterparts. These findings call into question the popularly held belief that villus tip cells are more susceptible to ischemia because of their location relative to the microvascular anatomy.

Animals↗
PubMed2002

Abundant expression of Dec1/stra13/sharp2 in colon carcinoma: its antagonizing role in serum deprivation-induced apoptosis and selective inhibition of procaspase activation.

The basic helix-loop-helix (bHLH) proteins are intimately associated with developmental events such as cell differentiation and lineage commitment. The HLH domain in the bHLH motif is responsible for dimerization, whereas the basic region mediates DNA binding. Based on sequence alignment and domain analysis, differentially expressed in chondrocytes/stimulated with retinoic acid/split and hairy-related proteins (DEC/STRA/SHARPs) represent a new class of bHLH proteins. The present study describes the functional characterization of DEC1. Subtractive experiments and blotting analyses demonstrated that DEC1 was highly expressed in colon carcinomas, but not in the adjacent normal tissues. Several cell cycle blockers markedly induced DEC1 expression. Stable transfectants with a tetracycline-inducible construct demonstrated that DEC1 caused proliferation inhibition, antagonized serum deprivation-induced apoptosis and selectively inhibited the activation of procaspases. These activities were highly correlated with the abundance of tetracycline-induced DEC1. Stable transfectants expressing a mutant DEC1 (lacking the DNA-binding domain) exhibited neither proliferation inhibition nor apoptotic antagonism, which suggests that DNA binding is required for these actions. Enzymic assays and immunoblotting analyses demonstrated that induction of DEC1 by tetracycline significantly decreased the activation of procaspases 3, 7 and 9 but not procaspase 8. The selective suppression on the activation of procaspases 3, 7 and 9 over procaspase 8 suggests that DEC1-mediated anti-apoptosis is achieved by blocking apoptotic pathways initiated via the mitochondria. The results functionally distinguish DEC1 from other bHLH proteins and directly link this factor to oncogenesis.

Adenocarcinoma↗
PubMed2005

Phenotypical enrichment strategies for microarray data analysis applied in a type II diabetes study.

Combining results from gene microarrays, clinical chemistry, and quantitative tissue histomorphology in an integrated bioinformatics setting enables prioritization of gene families as well as individual genes in a type II diabetes animal study. This new methodology takes advantage of a time-controlled mouse study as the animals progress from a normal phenotype to that of type II diabetes. Profiles from different levels of the biological hierarchy of unpooled entities provide an encompassing, system-wide view of biological changes. Here, phenotypic changes on the tissue-structural and physiological level are used as statistical covariants to enrich the gene expression analysis, suggesting correlative processes between gene expression and phenotype unlocked by multi-sample comparisons. We apply correlative and gene set enrichment procedures and compare the results to differential analysis to identify molecular markers. Evaluation based on ontological classifications proves changes in prioritization of disease-related genes that would have been overlooked by conventional gene expression analyses strategies.

Animals↗
PubMed1987

Dengue 4 virus monoclonal antibodies identify epitopes that mediate immune infection enhancement of dengue 2 viruses.

Nineteen monoclonal antibodies produced to dengue type 4 virus (DEN-4) strain 4328-S were tested for their ability to mediate antibody-dependent infection enhancement (ADE) with seven DEN-2 strains in P-388D1 mouse macrophage-like cells. In this first study of the distribution of enhancing epitopes on multiple DEN-2 strains reacted with monoclonal antibodies to a different serotype (DEN-4), DEN-4 monospecific antibodies produced ADE with DEN-2 viruses, indicating the presence of DEN-4-like determinants on DEN-2 viruses. Analysis differentiated at least one and possibly more DEN-2 strain subgroups, one of which (isolates AHF-110 and AHF-191) was previously identified by DEN-2 monoclonal antibody analysis. The study demonstrates the heterogeneous distribution of dengue complex and DEN-4 epitopes on DEN-2 strains. Monoclonal antibodies are valuable tools for study of the biology of ADE and its relation to dengue shock syndrome.

Animals↗
PubMed1994

Male mentally handicapped sex offenders.

BACKGROUND: Sex offences are overrepresented in the mentally handicapped, but information about the characteristics and offence behaviour of this group is limited. METHOD: A retrospective case note survey was made of 47 male patients referred for antisocial sexual behaviour. RESULTS: Of 191 offences and/or incidents committed, 55.5% were heterosexual, 24% indecent exposure, 12.4% homosexual, 13.6% serious and 3.6% involved physical assault. Average age of the offenders was 23.9 years, with a mean IQ of 59.5, and there was a high prevalence of family psychopathology, psychiatric illness, minor physical disabilities, sexual experience, impaired relationship skills and sexual recidivism. Recidivists showed a low specificity for offence type and age and sex of victim. Categorical analysis differentiated a 'sex offences only' group from a 'sex plus other offences' group in the key areas of psychopathology, offence behaviour and outcome; this was supported by factor analysis. CONCLUSION: The finding of two subgroups of mentally handicapped male sex offenders has important implications for prevention, assessment and treatment.

Adolescent↗
PubMed2000

Reversible programming of pluripotent cell differentiation.

We have undertaken an in vitro differentiation analysis of two related, interconvertible, pluripotent cell populations, ES and early primitive ectoderm-like (EPL) cells, which are most similar in morphology, gene expression, cytokine responsiveness and differentiation potential in vivo to ICM and early primitive ectoderm, respectively. Pluripotent cells were differentiated in vitro as aggregates (embryoid bodies) and the appearance and abundance of cell lineages were assessed by morphology and gene expression. Differentiation in EPL cell embryoid bodies recapitulated normal developmental progression in vivo, but was advanced in comparison to ES cell embryoid bodies, with the rapid establishment of late primitive ectoderm specific gene expression, and subsequent loss of pluripotent cell markers. Nascent mesoderm was formed earlier and more extensively in EPL cell embryoid bodies, and resulted in the appearance of terminally differentiated mesodermal cell types prior to and at higher levels than in ES cell embryoid bodies. Nascent mesoderm in EPL cell embryoid bodies was not specified but could be programmed to alternative fates by the addition of exogenous factors. EPL cells remained competent to form primitive endoderm even though this is not the normal fate of primitive ectoderm in vivo. The establishment of primitive ectoderm-like gene expression and inability to participate in embryogenesis following blastocyst injection is therefore not directly associated with restriction in the ability to form extra-embryonic lineages. However, the EPL cell embryoid body environment did not support differentiation of primitive endoderm to visceral endoderm, indicating the lack of an inductive signal for visceral endoderm formation deduced to originate from the pluripotent cells. Similarly, the inability of EPL cells to form neurons when differentiated as embryoid bodies was attributable to perturbation of the differentiation environment and loss of inductive signals rather than a restricted differentiation potential. Reversion of EPL cells to ES cells was accompanied by restoration of ES cell-like differentiation potential. These results demonstrate the ability of pluripotent cells to adopt developmentally distinct, stable cell states with altered differentiation potentials.

Animals↗
PubMed2005

Upregulation of class II beta-tubulin expression in differentiating keratinocytes.

The diverse functions of microtubules (MT) in different cells and tissues may be facilitated by compositional changes in tubulin isotypes. We obtained partial cDNA clones of class II beta-tubulin from a library of differentiating normal human epidermal keratinocytes (NHEK) cells, whereas screening via subtractive hybridization for genes involved in calcium-induced keratinocyte differentiation. Analysis of the isotypic composition of beta-tubulin from NHEK cells revealed elevations in class II beta-tubulin concentrations at both protein and message levels during cell differentiation, resulting in increased rates of incorporation of class II beta-tubulin into MT. Immunohistochemistry of normal and pathologic skin tissues showed that class II beta-tubulin occurred in the granular layer of the epidermis and in differentiated areas of carcinomas. Class II beta-tubulin was, however, not observed in the uppermost granular and cornified layers of normal epidermis. Further experiments showed that MT were likely to decay in the final stage of terminal differentiation during formation of the cornified envelope. Our results suggest that there is differential modulation of MT composition and stability during keratinocyte differentiation.

Acetylation↗
PubMed2000

Increased glyceraldehyde-3-phosphate dehydrogenase expression in renal cell carcinoma identified by RNA-based, arbitrarily primed polymerase chain reaction.

BACKGROUND: Renal cell carcinoma (RCC) comprises 85% of renal tumors and displays a great capacity to metastasize. The lack of diagnostic and prognostic markers complicates its early detection and in the majority of cases metastases are present at the time of diagnosis. METHODS: The current study reports on the identification of differentially expressed genes in RCC using random arbitrarily primed polymerase chain reaction (RAP-PCR). RESULTS: Four genes were identified, including glyceraldehyde-3-phosphate dehydrogenase (GAPDH), lactate dehydrogenase A (LDH A), human leukocyte antigen A (HLA A), and ferritin. GAPDH and HLA A were found to be overexpressed in 100% of the tumors and LDH A was increased in > 85% of the tumors analyzed compared with normal kidney counterparts. For GAPDH and LDH A higher protein levels in the tumors also were determined by Western blot analysis. Differential expression did not appear to correlate with gene amplification events as demonstrated by Southern blot analysis, indicating that regulatory mechanisms controlling the expression of these genes were altered. Finally, ferritin was judged to have a variable expression because it was decreased in approximately 50% of the tumors and augmented in 20%. The implications in proliferation and differentiation of all these genes were analyzed in RCC cell lines grown at different stages of confluency and additional information was obtained regarding expression of the GAPDH gene in proliferating primary cultures of normal and tumor cells derived from the same kidney samples. CONCLUSIONS: The authors conclude that RAP-PCR is a useful technique with which to identify rapidly differentially expressed genes in a given system. In addition, they also conclude that GAPDH is a potent marker of cell proliferation in kidney tumor cells whose overexpression appears to be a late event in the development of RCC.

Aged↗
PubMed2026

Dynamic Alterations in the Blood Transcriptome Characterize Drug Use Behavior and Co-Morbidities in Cocaine Use Disorder: A Preliminary Study.

Individuals with cocaine use disorder (CUD) who attempt abstinence experience craving and relapse that can benefit from multimodal treatment monitoring. Longitudinal studies linking behavioral manifestations in CUD to the blood transcriptome are not only limited but also computationally complex. Therefore, we developed an analytical pipeline to investigate the connection between drug use behaviors during abstinence and change in the blood transcriptome. We conducted a longitudinal study with CUD (n = 12 subjects) and collected behavioral metrics and blood RNA-seq at baseline, 3, 6, and 9 months. Our analytical pipeline of the high-dimensional data encompasses hierarchical k-means clustering to classify subjects to responder groups based on behavioral scores and abstinence duration, in silico cell deconvolution, differential analysis with correlated multivariate testing over time, gene set enrichment analysis, and gene co-expression with time splines and RNA-seq data. The pipeline captured dynamic changes in behavioral scores and abstinence duration in responder groups. Genes showing differential transcript-level expression were enriched in substance use and cardiovascular disease-associated genetic risk loci in responder groups. Lastly, time-dependent gene co-expression revealed dynamic changes related to immune processes, cell cycle, RNA-protein synthesis, and second messenger signaling for days of abstinence. This is a preliminary investigation, providing an innovative and scalable pipeline for blood-based longitudinal RNA-seq studies in CUD, potentially applicable to other substance use disorders. It outlines a data-driven approach for analyzing composite longitudinal drug use behavioral phenotypes with blood-based transcriptomics. We also demonstrate changes in drug use behaviors and the blood transcriptome during drug abstinence.

Humans↗
PubMedDate not supplied

[Investigations of humoral and cellular regulation mechanisms of the redox-homeostasis in cattle from ecological farming--2. Influence of race and age on the enzymatic redox control in the blood plasma].

The present study examined the effects of race and age on the redox potential behavior of blood plasma samples. The blood plasma from clinical healthy heifers (44) and calves (45) were investigated. The animals presented three different cattle races (Limousin, Angus and Hereford), grazed on pasture. The blood plasma test consists of two parts: analysis of the initial redoxpotential as well as the investigation of the action of biocatalists. ATP, ATP plus caffeine, GTP and FAD+ were used as an adding substances for the differential analysis. The influence of biocatalist on the redox balance and the reaction of the redox dependent enzymatic system of native plasma significant correlated with animal race and age.

Age Factors↗
PubMed1975

Thermodifferential analysis of ceramic implants.

Ceramic pelletes (in Al2O3 92% and in calcium aluminate) implanted into the muscles of rabbits for varying periods of time, and samples of similar material, not implanted but mixed with organic substances, have been analyzed by means of thermal differential analysis (TDA). Which hydrated calcium aluminates are formed from implants after contract with the tissue fluid, has been established. It has also been ascertained that the ceramic implants absorb organic substances from the tissue fluid, which, from a first evaluation, seem to be composed of simple molecules. The results of this initial inquiry have provided positive indications of the utility of TDA to assist in elucidating the phenomena that occur between implant and tissue.

Absorption↗
PubMed2006

[Evaluation on methodological problems in reports concerning quantitative analysis of syndrome differentiation of diabetes mellitus].

OBJECTIVE: To evaluate the quality of reports published in recent 10 years in China about quantitative analysis of syndrome differentiation for diabetes mellitus (DM) in order to explore the methodological problems in these reports and find possible solutions. METHODS: The main medical literature databases in China were searched. Thirty-one articles were included and evaluated by the principles of clinical epidemiology. RESULTS: There were many mistakes and deficiencies in these articles, such as clinical trial designs, diagnosis criteria for DM, standards of syndrome differentiation of DM, case inclusive and exclusive criteria, sample size and estimation, data comparability and statistical methods. CONCLUSION: It is necessary and important to improve the quality of reports concerning quantitative analysis of syndrome differentiation of DM in light of the principles of clinical epidemiology.

Bibliometrics↗
PubMed2004

Effect of growth factors on proliferation and phenotypic differentiation of human fetal neural stem cells.

Human fetal neural stem cells (hNSCs) can be expanded in vitro by mitogens or growth factors, such as basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and/or leukemia inhibitory factor (LIF). Their effects on proliferation rate and differentiation pattern of hNSCs, however, have not been fully characterized. In this study, we cultured hNSCs in seven regimens, including bFGF, EGF, and LIF, either alone or in combinations. Cells were maintained as neurospheres in treatment media for various periods, up to six passages. A combination of bFGF, EGF, and LIF expanded hNSCs more efficiently than any other treatment as determined by counting total cell numbers using a trypan blue exclusion assay, a WST-1 cell viability assay, and a bromodeoxyuridine incorporation flow cytometric analysis. Differentiation patterns of hNSCs expanded under different conditions were also analyzed. We reported previously that hNSCs primed in vitro with a combination of bFGF, heparin, and laminin (FHL) induced neuronal differentiation toward a cholinergic phenotype. In this study, we show that the FHL priming increases neuronal differentiation while decreasing astroglial generation in all treatment groups as determined by immunostaining. However, cells proliferated under different growth factor conditions do vary in their phenotypic differentiation patterns. Particularly, significant generation of cholinergic cells was observed only in hNSCs expanded with EGF/bFGF or EGF/bFGF/LIF, but not with other treatment regimens, even when they are exposed to the same priming procedure. Our results indicate that hNSCs are highly plastic, with their proliferation and differentiation potential dependent on different growth factor treatments.

Analysis of Variance↗
PubMed2004

Differences in expression of retinal pigment epithelium mRNA between normal canines.

A reference database of differences in mRNA expression in normal healthy canine retinal pigment epithelium (RPE) has been established. This database identifies non-informative differences in mRNA expression that can be used in screening canine RPE for mutations associated with clinical effects on vision. Complementary DNA (cDNA) pools were prepared from mRNA harvested from RPE, amplified by PCR, and used in a subtractive hybridization protocol (representational differential analysis) to identify differences in RPE mRNA expression between canines. The effect of relatedness of the test canines on the frequency of occurrence of differences was evaluated by using 2 unrelated canines for comparison with 2 female sibling canines of blue heeler/bull terrier lineage. Differentially expressed cDNA species were cloned, sequenced, and identified by comparison to public database entries. The most frequently observed differentially expressed sequence from the unrelated canine comparison was cDNA with 21 base pairs (bp) identical to the human epithelial membrane protein 1 gene (present in 8 of 20 clones). Different clones from the same-sex sibling RPE contained repetitions of several short sequence motifs including the human epithelial membrane protein 1 (4 of 25 clones). Other prevalent differences between sibling RPE included sequences similar to a chicken genetic marker sequence motif (5 of 25), and 6 clones with homology to porcine major histocompatibility loci. In addition to identifying several repetitively occurring, noninformative, differentially expressed RPE mRNA species, the findings confirm that fewer differences occurred between siblings, highlighting the importance of using closely related subjects in representational difference analysis studies.

Animals↗
PubMed2002

The scavenger receptor, cysteine-rich domain-containing molecule gp-340 is differentially regulated in epithelial cell lines by phorbol ester.

Gp-340 is a glycoprotein belonging to the scavenger receptor cysteine rich (SRCR) group B family. It binds to host immune components such as lung surfactant protein D (SP-D). Recent studies found that gp-340 interacts directly with pathogenic microorganisms and induces their aggregation, suggesting its involvement in innate immunity. In order to investigate further its potential immune functions in the appropriate cell lines, the expression of gp-340 in four conventional immune cell lines (U937, HL60, Jurkat, Raji), and two innate immune-related epithelial cell lines (A549 derived from lung and AGS from stomach), was examined by RT-PCR and immunohistochemistry. The resting immune cell lines showed weak or no gp-340 mRNA expression; while the two epithelial cell lines expressed gp-340 at much higher level, which was differentially regulated by phorbol myristate acetate (PMA) treatment. In the A549 cells, gp-340 was up-regulated along with the PMA-induced proinflammatory expression of both IL-6 and IL-8. In AGS cells, PMA down-regulation of gp-340 was seen in parallel with an up-regulation of the two mature gastric epithelial specific proteins TFF1 (trefoil factor 1) and TFF2, which are implicated as markers of terminal differentiation. Analysis of the distribution of gp-340, together with the TFFs and SP-D in normal lung and gastric mucosa, supported further our in vitro data. We conclude that the differential regulation of gp-340 in the two epithelial cell lines by PMA indicates that gp-340 s involvement in mucosal defence and growth of epithelial cells may vary at different body locations and during different stages of epithelial differentiation.

B-Lymphocytes↗
PubMed2002

Measurement of seeing and the atmospheric time constant by differential scintillations.

A simple differential analysis of stellar scintillations measured simultaneously with two apertures opens the possibility to estimate seeing. Moreover, some information on the vertical turbulence distribution can be obtained. A general expression for the differential scintillation index for apertures of arbitrary shape and for finite exposure time is derived, and its applications are studied. Correction for exposure time bias by use of the ratio of scintillation indices with and without time binning is studied. A bandpass-filtered scintillation in a small aperture (computed as the differential-exposure index) provides a reasonably good estimate of the atmospheric time constant for adaptive optics.

Journal Article↗
PubMed2002

[Sleep related breathing disorders in allergology practice].

1/3 of the contemporary population suffers from sleep disorders. Sleep related breathing disorders are very important from the clinical point of view, because they markedly contribute to high morbidity and mortality of this population. Pharyngeal and upper airway obstruction is the main cause of these disorders. Obstructive sleep apnoea (OSA) may be isolated or co-exist with different disorders of the respiratory or other systems. Prevalence of OSA is also reported in allergic diseases: allergic rhinitis, asthma and atopic dermatitis. The author presents and discusses 5 basis problems practically important from the point of view of the allergist, from the moment when his allergic patients complains of sleep disorders: 1. rough diagnosis of character of sleep and respiratory disorders; 2. their objective confirmation and severity classification; 3. etiology and differential analysis of causes; 4. patient education; 5. choice of individual therapeutic option.

Airway Obstruction↗