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PubMed1991

C-fos and c-jun proto-oncogene expression is decreased in psoriasis: an in situ quantitative analysis.

Psoriasis is a common, sometimes severe, non-malignant skin disease characterized by hyperproliferation and abnormal differentiation of keratinocytes. Because proto-oncogenes are implicated in both cell proliferation and differentiation, their expression could be modified in skin diseases such as psoriasis. The c-fos and c-jun proto-oncogenes, whose products associate to form a heterodimeric transcription factor, are among the first genes to be expressed when certain cells are stimulated to either proliferate or differentiate. Recent studies in our laboratory have shown that the c-fos proto-oncogene is highly expressed in normal human adult skin. In the present study, we used in situ hybridization with RNA to compare the expression and localization of c-fos and c-jun transcripts in 15 lesional and non-lesional psoriatic skin samples. Two clinical variants of psoriasis were studied: the most severe and chronic form or plaque-type psoriasis (N = 10) and rapidly resolutive guttate-type psoriasis (N = 5). Quantitative analysis was performed using a semi-automatic image analyzer and the "Starwise grain" software program. Our control samples included 10 normal skins and eight specimens from other benign hyperproliferative non-psoriatic skin diseases, consisting of three with inflammation (seborrheic dermatitis and atopic dermatitis), and 5 without inflammation (seborrheic keratoses). Control genes we used for in situ hybridization and RNA integrity were keratin 14, which is expressed in the epidermis and was normally expressed in all tissue analyzed, and ribosomal RNA. Our data showed that c-fos and c-jun were expressed to an equivalent extent, both spatially and quantitatively, in all specimens tested. Expression was significantly decreased in plaque-type but not in guttate-type psoriasis. It was also decreased in the three other benign inflammatory cutaneous hyperproliferative disorders, but not in the five non-inflammatory cases. These results were surprising because hyperproliferation was here associated with a decrease in proto-oncogene expression, thus suggesting that c-fos and c-jun do not play a crucial role in the control of keratinocyte proliferation in vivo. However, their reduced expression in some abnormally differentiated skins indicates that both c-fos and c-jun proto-oncogenes may play a key role in keratinocyte differentiation. Their altered expression correlated with severity of the disease and the presence of an inflammatory infiltrate. These data offer a new insight into the role and regulation of these proto-oncogenes in vivo in humans.

Chronic Disease↗
PubMed2002

[Colorectal exfoliated cell in stool and its nuclear DNA quantitative analysis for diagnosis of colorectal cancer].

BACKGROUND AND OBJECTIVES: Detection of colorectal exfoliated epithelial cells and their nuclear DNA content may provide another non-invasive way of screening and early diagnosis of colorectal cancer. This study was designed to analyze the roles of exfoliated cells in stool and its nuclear DNA content in diagnosis of colorectal cancer. METHODS: 1. One hundred and seventy nine individuals were selected, forty-six of them had pathological confirmation of colorectal carcinoma. The other 133 persons had no colonoscopic evidence of colorectal malignancy and therefore served as control. Exfoliated cells in the stool were isolated by elutriation, and the elutriation means was modified Iyengar's method. All individuals in the study had stool specimens for occult blood test(FOBT). 2. Nuclear DNA content and morphometric quantitative analysis in the exfoliated cells was performed on the 33 patients with colorectal cancer and 30 individuals served as control. The parameters selected in this study were DNA content, nuclear area, nuclear irregular index, and percent of > or = 5C cells. RESULTS: 1 Exfoliative cytology and FOBT: In 35 of 46 cases of colorectal malignancy(76.09%), cytology was positive: 5 cases demonstrated dysplastic cells, 4 cases indicated suspected carcinoma cells, 26 cases showed carcinoma cells. The positive rate of exfoliated cells had no significant relation to locations, sizes, histomorphologies, histological differentiations, Dukes stages, and lymph node metastases of the lesion(P > 0.05). Exfoliative cytology had a 98.50% (131/133) specificity for colorectal cancer in the study. The sensitivity for colorectal cancer was no significant difference between the two methods of exfoliative cytology and FOBT (76.09% vs 84.78%, P > 0.05), but the specificity for colorectal cancer, exfoliative cytology was significant higher than FOBT(98.50% vs 73.68%, P < 0.05). 2. DNA analysis of exfoliated cells nuclear, DNA content, nuclear area, nuclear irregular index and percent of > or = 5C cells in the stool were significant higher in colorectal cancer than in control group(P < 0.05). The percentage of > or = 5C cells were significantly associated with histological grade (P < 0.05). CONCLUSIONS: 1. Detection of exfoliated cells in stool plays an important role in diagnosis of colorectal cancer. Testing of FOBT and exfoliated cells sequentially hopes to be a new useful non-invasive test for screening of colorectal cancer. 2. DNA analysis of exfoliated cell in stool may provide an objective method of determining malignant grades and diagnosis of colorectal cancer.

Adult↗
PubMed1978

Quantitative analysis of spleen cell and immunoglobulin allotype composition of (CBAxCBA/H-T6) in equilibrium C57BL/6 allophenic mice.

Fifteen (CBAxCBA/H-T6) in equilibrium C57BL/6 allophenic mice were analyzed for the parental composition of their spleen white blood cells and serum immunoglobulin allotypes. The spleen compositions, assessed by a microcytotoxicity test, showed that the animals covered a range of parental cell mixtures. The allotype compositions of the IgG2a and IgG2b subclasses were determined by a solid phase radioimmunoassay. Although the absolute amount of IgG2a and IgG2b varied markedly from animal to animal, the percentage of the two allotypes in each subclass was remarkably constant. This suggests that the synthesis of IgG2a and IgG2b may be under a coordinate control system. Also, the spleen cell compositons were in excellent agreement with both the IgG2a and IgG2b subclass compositions. The quantitative analysis of the immune systems of allophenic mice may be first step toward the understanding of the mechanisms of tolerance of histoincompatible cells in allophenic mice.

Animals↗
PubMed1993

Determination of allele frequencies at loci with length polymorphism by quantitative analysis of DNA amplified from pooled samples.

We present a new method that allows rapid determination of allele frequencies at loci exhibiting length polymorphism. In this method a fluorescence-labeled PCR primer is used to amplify the polymorphic region from pooled DNA samples originating from a large number of individuals. The fluorescent PCR products are separated by gel electrophoresis on an automatic DNA sequencer and the relative amount of the PCR products are determined. The distribution of the PCR products obtained from the alleles present in the pooled samples directly corresponds to the allele frequency in the population in question. The allele frequencies at a short tandem repeat locus in the von Willebrand factor gene and at the D1S80 locus were determined in the Finnish population. We found that the allele frequencies determined by quantitative analysis of PCR products from pooled DNA samples and by analyzing individual samples were in good agreement.

Alleles↗
PubMed2002

Isolation and quantitative analysis of phenolic antioxidants, free sugars, and polyols from mango (Mangifera indica L.) stem bark aqueous decoction used in Cuba as a nutritional supplement.

An aqueous decoction of mango (Mangifera indica L.) stem bark has been developed in Cuba on an industrial scale to be used as a nutritional supplement, cosmetic, and phytomedicine. Previously we reported its antioxidant activity, and we concluded that the product could be useful to prevent the production of reactive oxygen species and oxidative tissue damage in vivo. A phytochemical investigation of mango stem bark extract has led to the isolation of seven phenolic constituents: gallic acid, 3,4-dihydroxy benzoic acid, gallic acid methyl ester, gallic acid propyl ester, mangiferin, (+)-catechin, (-)-epicatechin, and benzoic acid and benzoic acid propyl ester. All structures were elucidated by ES-MS and NMR spectroscopic methods. Quantitative analysis of the compounds has been performed by HPLC, and mangiferin was found to be the predominant component. Total polyphenols were assayed also by the Folin-Ciocalteu method. The free sugars and polyols content was also determined by GC-MS.

Antioxidants↗
PubMed2003

Automated quantitative analysis of nocturnal jejunal motor activity identifies abnormalities in individuals and subgroups of patients with slow transit constipation.

OBJECTIVE: Small bowel dysmotility has previously been demonstrated in some patients with slow transit constipation (STC), suggesting a generalized intestinal disorder. However, no study has addressed whether the incidence of small intestinal dysfunction differs between subgroups of patients in this heterogeneous population. Using appropriate methodology, we aimed to determine prospectively the proportion of individuals with abnormal small bowel motility, and to assess whether heterogeneity in terms of pattern of colonic transit delay (based on (111)In diethylene-triamine-pentaacetic acid (DTPA) isotope scintigraphy), or mode of onset (based on clinical history) is of importance. METHODS: Thirty-seven patients with STC underwent 24-h ambulatory jejunal manometry; data were compared with those obtained in 38 healthy controls. Automated quantitative analysis of seven variables of the nocturnal migrating motor complex was performed, to assess whether differences existed between groups, and whether individual patients had evidence of small intestinal dysmotility, defined as two or more measures of migrating motor complex variables outside the normal range. Four variables differed significantly between STC patients and controls: in phase III, propagation was slower, duration was longer, and contraction amplitude was higher; in phase II, contraction frequency was increased. Seven of 24 patients with a generalized pattern of colonic transit delay had abnormal small bowel motility compared with none of 13 with a left-sided delay (p < 0.04). These included four patients with chronic idiopathic symptoms and three with acquired symptoms. Approximately one third of patients with a generalized delay in colonic transit had evidence of jejunal enteric neuromuscular dysfunction. Individual patients with a left-sided colonic delay did not satisfy the criteria for nocturnal small bowel dysmotility, but as a group, some differences were noted from controls. In contrast to previous reports, evidence of generalized enteric dysmotility may be present irrespective of the mode of onset.

Adolescent↗
PubMed1996

[Quality evaluation on Paeoniae radix. I. Quantitative analysis of monoterpene glycosides constituents of Paeoniae radix by means of high performance liquid chromatography. Comparative characterization of the external figures, processing method and the cultivated areas].

The constituents of monoterpene in Paeoniae Radix were analyzed by means of high performance liquid chromatography (HPLC) using hitherto elucidated eight monoterpene glycosides, paeoniflorin 1, oxypaeoniflorin 2, benzoylpaeoniflorin, 3, benzoyloxypaeoniflorin 4, galloylpaeniflorin 5, galloyloxypaeoniflorin 6, albiflorin 7 and lactiflorin 8 and a monoterpene paeoniflorigenone 9. In sixty seven kinds of Paeoniae Radix collected in Japan, China, and South and North Korea those monoterpene constituents 1-9 were found to exist in the amounts of ranging 0.12-9.61% 1, 0.06-10.8% 2, 0.02-0.79% 3, 0.24-0.47% 4, 0.25-2.53% 5, 0.05-2.86% 6, 0.09-2.76% 7, 0.35-0.64% 8 and 0.01-0.49% 9 respectively. In addition, these results obtained by the quantitative analysis were discussed from the viewpoint of the external figures and processing methods.

China↗
PubMed2006

Quantitative analysis of DNA-protein interactions using double-labeled native gel electrophoresis and fluorescence-based imaging.

We have developed a sensitive, non-radioactive method to assess the interaction of transcription factors/DNA-binding proteins with DNA. We have modified the traditional radiolabeled DNA gel mobility shift assay to incorporate a DNA probe end-labeled with a Texas-red fluorophore and a DNA-binding protein tagged with the green fluorescent protein to monitor precisely DNA-protein complexation by native gel electrophoresis. We have applied this method to the DNA-binding proteins telomere release factor-1 and the sex-determining region-Y, demonstrating that the method is sensitive (able to detect 100 fmol of fluorescently labeled DNA), permits direct visualization of both the DNA probe and the DNA-binding protein, and enables quantitative analysis of DNA and protein complexation, and thereby an estimation of the stoichiometry of protein-DNA binding.

Animals↗
PubMed2001

Quantitative analysis from CT is prognostic for local control of supraglottic carcinoma.

BACKGROUND: The purpose of this study was to determine whether pretreatment imaging with CT was prognostic for control of the primary site in patients with squamous cell carcinoma of the supraglottic larynx. METHODS: Pretreatment CT studies were obtained on 28 patients treated definitively with radiation therapy for supraglottic larynx cancer between 1991 and 1997. Follow-up ranged from 20-58 months. RESULTS: Local control was achieved in 61% of patients. Tumor volumes ranged from 0-68.6 cm(3), with a median of 3.1 cm(3). Local control rates for tumors with volumes greater than or less than 8 cm(3) were 20% and 70%, respectively (p =.0077). Mean tumor volumes for patients with and without recurrences were 10 cm(3) and 3.4 cm(3), respectively. CONCLUSIONS: This study demonstrates that quantitative analysis from CT imaging is prognostic for control of the primary site when radiation therapy is given for treatment of supraglottic cancer.

Carcinoma, Squamous Cell↗
PubMed1991

A quantitative analysis of transparency in the human sclera and cornea using Fourier methods.

Cellular microstructure observed by scanning transmission electron microscopy (STEM) was analysed using Fourier methods. Fourier components of the density fluctuations in the sclera and the cornea of the human eye were quantified. The results show that the Fourier components responsible for the opacity of the sclera have sizes of the order of the wavelength of visible light. In the sclera, approximately 54% of the spectral energy of the density fluctuation falls in the range of 200-1100 nm (scattering range). In the cornea, approximately 24% of the total spectral energy falls in this range. The predominant Fourier components of the density fluctuations in the opaque sclera are approximately 300 nm in wavelength, whereas those of the transparent cornea are approximately 80 nm in wavelength. This method will be useful in quantitative analysis of microstructural changes accompanying biological phenomena such as normal development of transparency in the human lens, and abnormal loss of transparency during cataract formation.

Cornea↗
PubMed1998

Quantitative analysis of liver peroxisomes in rats intoxicated with peroxisomicine-A1.

Peroxisomes are single-membrane-bound organelles present in almost all eukaryotic cells. Hypolipidemic agents such as clofibrate, herbicides and plasticizers induce an increase in the number and size of peroxisomes from mammalian cells. However, there is no evidence of drugs causing a decrease in the number of these organelles. In this paper, we report the effect in vivo of toxin T-514 extracted from the plant Karwinskia humboldtiana, now re-named peroxisomicine-A1, on hepatic peroxisomes from rats intoxicated with this compound. Rats were treated with a single dose of 25 mg/kg of peroxisomicine-A1 and at different times were killed by decapitation. For the peroxisomal counting, liver tissue sections from control and treated rats were processed for the localization of catalase in peroxisomes. The results of the quantitative analysis demonstrated a significant decrease in the number of liver peroxisomes from rats intoxicated with peroxisomicine-A1. This finding suggests that peroxisomicine-A1 as in yeast, causes a damage to mammalian peroxisomes. The diminution in the number of peroxisomes could be a consequence of damage to the organelle, which is further removed by an autophagic process.

Animals↗
PubMed2002

Localization of phosphatidylinositol 4,5-P(2) important in exocytosis and a quantitative analysis of chromaffin granule motion adjacent to the plasma membrane.

A slow ATP-dependent priming step precedes a rapid, Ca(2+)-dependent triggering step in exocytosis in chromaffin cells and in most, if not all, differentiated secretory cells. A major component of ATP-dependent secretion in permeabilized cells reflects the maintenance of the polyphosphoinositides, especially PtdIns-4,5-P2. Here we summarize recent experiments with PH-GFP (binds to PtdIns-4,5-P2) that indicate that PtdIns-4,5-P2 is localized primarily on the plasma membrane in chromaffin cells, and that it is this pool that plays a role in exocytosis. It is demonstrated that transiently expressed PH-GFP inhibits secretion in subsequently permeabilized cells. Recent studies using total internal reflection fluorescent microscopy (TIRFM) to measure chromaffin granule motion adjacent to the plasma membrane are also summarized. The quantitative analysis indicates that chromaffin granule motion is highly restricted and suggests that chromaffin granules are caged or tethered immediately adjacent to the plasma membrane.

Adenosine Triphosphate↗
PubMed1993

Quantitative analysis of iris translucency in Fuchs' heterochromic cyclitis.

PURPOSE: To measure intraocular stray light and to quantify translucency of the iris and the surrounding ocular wall in both eyes of patients with Fuchs' heterochromic cyclitis; to investigate whether differences in paired eyes could be measured in patients without heterochromia or with a minimal degree of iris atrophy. METHODS: Intraocular stray light was measured by means of the direct compensation technique. A modification of this technique was used to quantify translucency of the iris and the surrounding ocular wall. RESULTS: Intraocular stray light proved to be significantly higher in all patients with Fuchs' heterochromic cyclitis (both eyes) than in normal control subjects. Translucency of the iris and the ocular wall around it were increased in the patients with Fuchs' heterochromic cyclitis, including patients without heterochromia or with a minimal degree of iris atrophy. CONCLUSIONS: Quantitative analysis of translucency was used to determine iris depigmentation in vivo in patients with Fuchs' heterochromic cyclitis. This technique showed that the process of atrophy and depigmentation in patients with Fuchs' heterochromic cyclitis is probably not restricted to the iris, but also occurs in the surrounding ocular wall. More studies, including other uveitis groups, are necessary to investigate if this technique can be used as a diagnostic tool in Fuchs' heterochromic cyclitis.

Adult↗
PubMed1982

A quantitative analysis of leptomeric fibrils in an adriamycin/carnitine chronic mouse model.

A quantitative ultrastructural analysis of leptomeres in cardiac muscle was performed using an adriamycin/carnitine chronic mouse model. Five animals from each of the following four groups were sacrificed for evaluation by electron microscopy: the control group received saline injections only; the carnitine group received carnitine orally; the adriamycin group received a chronic dose of adriamycin (10 mg/kg over 4 weeks); the adriamycin/carnitine group received both the adriamycin and carnitine dosage regimens. The leptomeres were quantitated using a double-blind method. The total number of leptomeres were scored for 20 random low-power electron micrographs from each of 5 animals within each group. The adriamycin group contained 3 of the 4 lowest leptomere scores. The relationships between myofibrillar disruption (MD) and the number of leptomeres found in an intramyofibrillar location(IML) were then determined for each of the four groups using linear regression analysis. In the control group, increased MD was associated with a decrease in the number of IML. In the carnitine group, increased MD was associated with an increase in IML. In the adriamycin group, no significant change in the number of IML occurred with increasing levels of MD. The adriamycin/carnitine group was similar to the control group. It was concluded from this study that 1) a relationship exists between MD and the number of IML in normal murine cardiac muscle; 2) carnitine produces an inverse effect on this normal relationship; 3) adriamycin disturbs the normal relationship between IML and MD; and 4) the administration of carnitine to adriamycin-treated animals restored the normal relationship. The physiological significance of the leptomere to the cardiac cell is then discussed.

Animals↗
PubMed1997

Efficacy of quantitative analysis of Epstein-Barr virus-infected peripheral blood lymphocytes by in situ hybridization of EBER1 after living-related liver transplantation: a case report.

BACKGROUND: We describe a 1-year-old female who underwent living-related liver transplantation for biliary atresia and developed Epstein-Barr virus (EBV)-related posttransplant lymphoproliferative disorder. This disorder was resolved after withdrawal of immunosuppression therapy and administration of a high dose of acyclovir. METHODS: To quantify the extent of EBV activation and EBV load in peripheral blood, we measured the levels of EBV-infected peripheral lymphocytes by in situ hybridization (ISH) of EBV-encoded small mRNA 1 (EBER1). RESULTS: The decline in the number of EBER1-positive lymphocytes (from 362/50,000 mononuclear cells to 0/50,000) after treatment was in accord with the patient's clinical improvement. CONCLUSIONS: This finding showed that quantitative analysis of EBV-infected peripheral lymphocytes by ISH of EBER1 is very useful for monitoring the EBV load and response to treatment of patients with EBV-related disorders. Furthermore, ISH may become an important tool for the early diagnosis and prevention of life-threatening posttransplant lymphoproliferative disorder in posttransplant patients.

Acyclovir↗
PubMed2007

Can anti-migratory drugs be screened in vitro? A review of 2D and 3D assays for the quantitative analysis of cell migration.

The aim of the present review is to detail and analyze the pros and cons of in vitro tests available to quantify the anti-migratory effects of anti-cancer drugs for their eventual use in combating the dispersal of tumor cells, a clinical need which currently remains unsatisfied. We therefore briefly sum up why anti-migratory drugs constitute a promising approach in oncology while at the same time emphasizing that migrating cancer cells are resistant to apoptosis. To analyze the pros and cons of the various in vitro tests under review we also briefly sum up the molecular and cellular stages of cancer cell migration, an approach that enables us to argue both that no single in vitro test is sufficient to characterize the anti-migratory potential of a drug and that standardization is needed for the efficient quantitative analysis of cell locomotion in a 3D environment. Before concluding our review we devote the final two parts (i) to the description of new prototypes which, in the near future, could enter the screening process with a view to identifying novel anti-migratory compounds, and (ii) to the anti-migratory compounds currently developed against cancer, with particular emphasis on how these compounds were selected before entering the clinical trial phase.

Animals↗
PubMed1989

[Quantitative analysis of chromosomal localization of two human cloned satellite DNA III sequences by in situ hybridization].

Chromosomal location of two cloned human satellite DNA III sequences pPD9 and pPD18 has been studied in 30 individuals by in situ hybridization. Pericentromeric localization of the DNA subsets studied was found in practically all chromosomes of the set. The majority of label was observed over the pericentromeric region of chromosome 9 (38.3% for pPD18 clone and 26.2% for pPD9), the short arm of chromosome 15 (17.2% - the pPD9 clone and 10.6% - the pPD18 clone) and the distal part of the long arm of Y chromosome (19.6% - the pPD9 clone and 15.4% - the pPD18 clone). Besides significant interchromosomal differences, moderately pronounced interindividual differences were also detected in the number of grains over the regular sites of the chromosomal location. Pretreatment of slides with DA/DAPI induced differences in the results of quantitative analysis is described.

Base Sequence↗
PubMed1989

Quantitative analysis of the number and distribution of neurons richly innervated by GABA-immunoreactive axons in the rat superior cervical ganglion.

The superior cervical ganglion of rats contains a considerable number of nerve fibers with GABA-like immunoreactivity which show a nonuniform distribution within the ganglion. The topography of these fibers has been analyzed by using antibodies raised against GABA-BSA-glutaraldehyde complexes. GABA-positive axons and axon varicosities accumulated around a subpopulation of principal ganglion cells forming basketlike patterns. These neurons richly innervated by GABA-positive axons (RIG-neurons) in turn were aggregated in patches with strong immunoreactivity. The size and packing density of the patches containing RIG-neurons and GABA-positive axons approaching them had rostral-to-caudal and medial-to-lateral gradients. Similar patterns were found in right and left ganglia. In five ganglia, a quantitative analysis revealed on average 1,344 RIG-neurons per ganglion representing about 5% of the total neuron population, with small variations (standard deviation 122) despite the highly variable shape of the ganglia. The distribution of RIG-neurons resembles that of neurons sending their axons into the internal carotid nerve. To check this possible correlation, HRP was injected into the eye and applied to the transected external carotid nerve. Double staining for the retrogradely transported peroxidase and GABA immunohistochemistry revealed that RIG-neurons formed a small subpopulation of retrogradely labelled neurons in both experiments. This suggests that RIG-neurons innervate various target organs. This conclusion is in agreement with the observation that RIG-neurons also exist in other sympathetic ganglia. Data presented suggest that sympathetic ganglion cells can be classified on the basis of non-uniform innervation patterns formed by axons that use different neurotransmitters.

Animals↗