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PubMed2005

Influence of CYP2C19 polymorphism and Helicobacter pylori genotype determined from gastric tissue samples on response to triple therapy for H pylori infection.

BACKGROUND & AIMS: The relationship between single nucleotide polymorphisms (SNPs) and clinical outcomes has been intensively studied. We intended to determine SNPs of CYP2C19 and 23S rRNA of Helicobacter pylori by using rapid urease test (RUT)-positive gastric mucosal samples. METHODS: One hundred thirty-nine patients with H pylori -positive results based on RUT completed 1-week treatment with lansoprazole 30 mg twice a day, clarithromycin 200 mg 3 times daily, and amoxicillin 500 mg 3 times daily. SNPs from adenine to guanine at positions 2142 and 2143 of 23S rRNA of H pylori (A2142G and A2143G) and SNPs from guanine to adenine at positions 681 in exon 5 (* 2 ) and 636 in exon 4 (* 3 ) of CYP2C19 were determined by the serial invasive signal amplification reaction assay by using DNAs extracted from gastric tissue samples already used for RUT. Minimum inhibitory concentrations of clarithromycin for H pylori were determined by culture test. CYP2C19 genotypes were classified into the rapid metabolizer (* 1 /* 1 ), intermediate metabolizer (* 1 /* 2 or * 1 /* 3 ), and poor metabolizer (* 2 /* 2 , * 2 /* 3 , or * 3 /* 3 ) groups. RESULTS: H pylori strains with A2142G or A2143G mutation had higher minimum inhibitory concentrations for clarithromycin. Cure rates in rapid, intermediate, and poor metabolizer groups were 57.8% (95% confidence interval, 42.1%-72.4%), 88.2% (78.1%-94.8%), and 92.3% (74.9%-99.1%), respectively ( P < .001). Cure rates in strains with and without A2142G or A2143G mutation were 48.3% (29.4%-67.5%) and 87.3% (79.5%-92.7%), respectively ( P < .001). CONCLUSIONS: SNPs of CYP2C19 and 23S rRNA of H pylori using RUT-positive gastric mucosal samples could be predictable determinants for H pylori eradication by triple therapy.

2-Pyridinylmethylsulfinylbenzimidazoles↗
PubMed1967

Antigen-induced changes in lymphoid cell histones. IV. Changes in solubility of isolated chromatin.

In this study we have examined the solubility of deoxyribonucleoprotein (DNP) isolated from control and antigen-affected thymocytes. 2-M sodium chloride extracts containing the DNP of rat thymus glands were serially diluted. A comparison was made of the effect of dilution on fiber formation in the control and test series. Fiber formation is usually complete for the control material at a salt concentration between 0.63 and 0.57 M. The test material shows some fiber formation within this range. However, a significant portion of the DNP is precipitated at dilutions of 0.54-0.48 M. Ammoniacal silver (A-S) stains the control fibers a characteristic yellowish color. With the test material, those fibers formed within the control range tended to be stained yellowish brown by A-S, whereas those formed only after greater dilution stained blackish. These data, coupled with our previous observations on altered A-S staining, clearly demonstrate an antigen-induced physical and/or chemical alteration of the histone or histone-DNA complex of lymphoid cell chromatin.

Animals↗
PubMed2001

Demonstration of focal p53 expression without genetic alterations in endometriotic lesions.

Their monoclonal origin (as indicated by recent investigations) indicates the neoplastic nature of most endometriotic lesions. p53, a representative tumor suppressor, regulates cell proliferation, and genetic alterations in p53 are involved in carcinogenesis in a wide variety of human cancers. The aim of this study was to examine endometriotic lesions for p53 expression and genetic alterations in p53. An immunohistochemical study revealed that 20% (13/64) of endometriotic lesions showed focal p53 expression in the epithelial cells. Using serial paraffin sections, we employed a microdissection method to extract DNA from the endometriotic tissues that showed p53 expression. No mutations were found in exons 5-8 in p53 by cleavase fragment length polymorphism scanning and polymerase chain reaction-DNA sequencing. Moreover, neither loss of heterozygosity nor microsatellite instability was detected at the microsatellite marker sites of p53. These results suggest that the focal p53 expression recognized in the endometriotic epithelia may be due to overproduction of wild-type p53 protein.

Adult↗
PubMed2004

Internal apical resorption and its correlation with the type of apical lesion.

AIM: To determine the presence of various periapical pathologies and their association with the presence and extent of internal apical inflammatory root resorption in human teeth. METHODOLOGY: A total of 75 root apices from extracted teeth with periapical lesions were examined. Semi-serial sections of soft tissue lesions were stained with HE. The lesions were classified as non-cystic or cystic, and according to the degree of abscess severity: 0, 1, 2 or 3. The apices were reduced to 3 mm in length and longitudinally cut so that the internal aspect could be analyzed under scanning electron microscopy (SEM). Internal root resorption was also classified as 0, 1, 2 or 3 according to the extent of the resorbed area. Additionally, six vital teeth were used as a control. RESULTS: Non-cystic lesions with severe abscesses were the most common finding (70.7%), while 20% of the lesions were cystic (4% little or no abscess; 16% severe abscesses). Non-cystic lesions with little or no abscess comprised 9.3% of the sample. Of the root canals containing periapical lesions, 48% had internal apical resorption in more than half of the area, while 25.3% of the sample had no internal resorption. Resorption degree 1 was identified in 12% of the cases, and 14.7% showed resorption degree 2. The control group displayed significantly less internal resorption than the test groups. CONCLUSIONS: Most periapical lesions (86.7%), whether cystic (16.0%) or non-cystic (70.7%), showed large collections of acute inflammatory cells. Apical internal resorption was present in 74.7% of roots and was associated with periapical lesions. There was no correlation between internal apical resorption and the histological diagnosis of the lesions.

Analysis of Variance↗
PubMed2004

Plasticity of mouse renal collecting duct in response to potassium depletion.

Plasticity of mouse renal collecting duct in response to potassium depletion.--Renal collecting ducts are the main sites for regulation of whole body potassium balance. Changes in dietary intake of potassium induce pleiotropic adaptations of collecting duct cells, which include alterations of ion and water transport properties along with an hypertrophic response. To study the pleiotropic adaptation of the outer medullary collecting duct (OMCD) to dietary potassium depletion, we combined functional studies of renal function (ion, water, and acid/base handling), analysis of OMCD hypertrophy (electron microscopy) and hyperplasia (PCNA labeling), and large scale analysis of gene expression (transcriptome analysis). The transcriptome of OMCD was compared in mice fed either a normal or a potassium-depleted diet for 3 days using serial analysis of gene expression (SAGE) adapted for downsized extracts. SAGE is based on the generation of transcript-specific tag libraries. Approximately 20,000 tags corresponding to 10,000 different molecular species were sequenced in each library. Among the 186 tags differentially expressed (P < 0.05) between the two libraries, 120 were overexpressed and 66 were downregulated. The SAGE expression profile obtained in the control library was representative of different functional classes of proteins and of the two cell types (principal and alpha-intercalated cells) constituting the OMCD. Combined with gene expression analysis, results of functional and morphological studies allowed us to identify candidate genes for distinct physiological processes modified by potassium depletion: sodium, potassium, and water handling, hyperplasia and hypertrophy. Finally, comparison of mouse and human OMCD transcriptomes allowed us to address the question of the relevance of the mouse as a model for human physiology and pathophysiology.

Acid-Base Equilibrium↗
PubMed1978

Assessment of regional myocardial blood flow and regional fractional oxygen extraction in dogs, using 15O-water and 15O-hemoglobin.

A new approach to the assessment of regional myocardial blood flow and fractional oxygen extraction has been developed using 15O-water (H2-15O) and 15O-hemoglobin (15O-Hb). Bolus doses (1 mCi) of H2-15O and 15O-Hb were injected 10 minutes apart into the left main coronary artery of 12 normal dogs. Sequential images of regional myocardial tracer clearance were obtained over 5 minutes with a positron camera. Myocardial blood flow calculated from the monoexponential washout of H2-15O after background correction was 78 +/- 6 (SE) ml/100 g per min. Functional images of regional blood flow in which the image of peak activity was divided by the integrated image of H2-15O washout were derived by computer processing. These images demonstrated homogeneous blood flow in the normal myocardium. Fractional myocardial O2 extraction was determined from an image of initial distribution of O2 used (obtained by extrapolating back to time zero the series of images obtained after 15O-Hb administration), divided by initial distribution of O2 delivered (obtained by back extrapolating H2-15O washout). These functional images showed uniform distribution of fractional O2 extraction in the normal myocardium. Thus, these studies show that regional myocardial blood flow and regional oxygen extraction can be measured simultaneously by sequential imaging after serial intracoronary injections of H2-15O and 15O-Hb.

Animals↗
PubMed1982

Characterization of gamma-melanotropin-like immunoreactivity and its secretion in an adrenocorticotropin-producing mouse pituitary tumor cell line.

gamma MSH, a putative hormone in the N-terminal region of the ACTH/beta-endorphin (beta-EP) precursor protein, was studied by RIA with an antiserum against gamma 3MSH in ACTH-producing mouse pituitary tumor cells, AtT-20/D16v. Serial dilution of the culture medium or the cell extract gave parallel lines to the standard curve in the RIA for gamma MSH. Rat median eminence extracts enhanced the release of gamma MSH-like immunoreactivity (gamma MSH-LI) concomitant with ACTH-like immunoreactivity (ACTH-LI) and beta-EP-like immunoreactivity (beta-EP-LI). Dexamethasone suppressed the release of gamma MSH-LI as well as ACTH-LI and beta-EP-LI. Gel exclusion chromatography of the culture medium and the cell extract has revealed that gamma MSH-LI consists of two peaks; one eluted near the elution position of beta-lipotropin and the other near the elution position of beta-EP. There was no peak corresponding to the elution position of synthetic gamma 3MSH. However, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) has demonstrated that gamma MSH-LI migrated at five positions with molecular weights of 31K, 21-23K, 16-17K, 13-14K, and 3.8K, respectively. The 31K gamma MSH coincided with the migration position of 31K ACTH of 31K beta-EP, and 21-23K gamma MSH coincided with the position of 21-23K ACTH on SDS-PAGE. The 16-17K gamma MSH coincided with the mouse 16K fragment (reported by Eipper and Mains) of ACTH-beta-lipotropin precursor protein in the migration in SDS-PAGE and in immunoreactivity to anti-gamma MSH antiserum. [3H]Glucosamine was incorporated into 16K, 13K, and 3.8K gamma MSH. These results suggest that AtT-20/D16v cells produce gamma MSH-LIs with molecular weights of 31K, 21-23K, 16-17K, 13-14K, and 3.8K, and they are secreted concomitantly with ACTH-LI and beta-EP-LI.

Adrenocorticotropic Hormone↗
PubMed1992

Suppression of epicutaneous reactivity by terfenadine and loratadine.

We compared the relative antihistaminic effects of loratadine (10 mg), terfenadine (120 mg), and placebo in patients with tree pollen-induced allergic rhinitis. Wheals were produced by epicutaneous tests with serial dilutions of histamine phosphate, tree and grass pollen extracts before ingestion of medication. Repeat tests were performed after seven days of treatment. Terfenadine was more effective than both loratadine (P less than .01) and placebo (P less than .001) at suppressing histamine induced wheals. Terfenadine was more effective than placebo (P less than .01), but not loratadine at inhibiting both tree-induced and grass-induced epicutaneous reactions. In this study, terfenadine was more potent than loratadine in inhibiting both histamine-induced and allergen-induced epicutaneous wheals.

Cyproheptadine↗
PubMed1992

[Detection of mycobacteria by polymerase chain reaction].

The polymerase chain reaction (PCR) was used to detect mycobacterial DNA sequences in the cultured or the clinical specimens. Four oligonucleotide primers derived from the sequence of a gene coding 65-kilodalton antigen of Mycobacterium tuberculosis amplified DNA samples of all the 11 species of mycobacteria tested. Serial dilution of M. bovis BCG showed that DNA extracted from only 12 bacilli was enough for the detection by PCR method. However, mycobacteria in sputum were detected by PCR when more than 10(3) bacilli were present. The PCR method may become a useful tool for the rapid diagnosis of mycobacterial infections.

Base Sequence↗
PubMed1985

Surgically induced corneal astigmatism and its spontaneous regression.

Serial keratometry readings were obtained pre- and postoperative cataract extraction utilizing all silk closure. The induction of with-the-rule cylinder and its spontaneous regression was plotted over 24-weeks. The data are compared to others utilizing all nylon closure.

Astigmatism↗
PubMedDate not supplied

Mild dental findings associated with severe osteogenesis imperfecta due to a point mutation in the alpha 2(I) collagen gene demonstrate different expression of the genetic defect in bone and teeth.

Serial intraoral photographs, radiographs, and ground sections from an extracted upper permanent canine served to characterize dental abnormalities in a 15-year-old girl suffering from severe (type III) osteogenesis imperfecta (OI) due to a point mutation that substituted glycine 688 of the alpha 2(I) chain of collagen I by serine. Dental records showed that all deciduous teeth exhibited clinical and radiographic characteristics of dentinogenesis imperfecta (DI), whereas permanent teeth including the removed canine appeared normal, although pulp chambers contained unusually large denticles. Despite the unconspicuous clinical appearance of the canine, histologic sections revealed small, canal-like, hypomineralized hard tissue patches that lacked a regular tubular structure and occupied a narrow band of the bulk of normal circumpulpal dentin at about the level of the cemento-enamel junction. The finding that a mutation in the gene for the alpha 2(I) collagen chain with serious consequences in bone has only minor effects in teeth would suggest that odontoblasts, unlike osteoblasts, can largely compensate for this particular genetic defect, possibly by excluding the abnormal alpha 2(I) chains and forming alpha 1(I) homotrimeric collagen I. The discrepant consequences in deciduous as opposed to permanent teeth and the specific localization of the dentinal abnormalities in permanent teeth lead us to speculate that the exclusion of defective alpha 2(I) chains could depend on the developmental stage and/or the rate of extracellular matrix formation.

Adolescent↗
PubMed1999

Overexpression of tissue-type plasminogen activator in atherosclerotic human coronary arteries.

UNLABELLED: The plasminogen activator (PA)/plasmin system is involved in various pathological processes that are considered important features of atherogenesis and atherothrombosis. These include the proteolysis of fibrin deposits and extracellular matrix components as well as the induction of cell migration and mitogenesis. Tissue-type PA (TPA) is a key enzyme mediating plasminogen to plasmin conversion. TPA plasma concentrations are elevated in patients with advanced atherosclerosis and correlate with an increased risk for myocardial infarction and stroke. In this study, we have analysed the content and expression of TPA in human coronary arteries and their relation to the presence and severity of atherosclerotic lesions. METHODS: Segments of coronary arteries obtained from heart explants (n = 15) were classified by the presence and types of atherosclerotic lesions. TPA was quantitatively determined in protein extracts of intimal and medial layers. In situ hybridization and immunohistochemical analyses were performed on serial sections of representative tissue specimens. RESULTS: PA activity entirely attributable to the presence of active TPA was consistently detected in the protein extracts. Extractable TPA antigen and activity showed a significant graded increase in relation to the presence and severity of atherosclerotic lesions. The ratios of active over total TPA were increased several-fold in extracts of advanced lesions despite a concomitant threefold increase in TPA complexed to its inhibitor PA-1. In macroscopically normal arterial segments and in early lesions, TPA was expressed in the endothelium and in colocalization with vascular smooth muscle cells (VSMCs). In advanced plaques, TPA mRNA was mainly detected in the lateral regions of the fibrous caps in association with migrating VSMCs and in the vicinity of the core areas infiltrated by CD68-positive macrophages. CONCLUSIONS: TPA content and expression is consistently increased in relation to the severity of the lesions in atherosclerotic coronary arteries. This may contribute to plaque destabilization and disruption. Conversely, the increased intramural TPA activity may counteract mural fibrin deposition.

Arteries↗
PubMed1992

Extract of kava (Piper methysticum) and its methysticin constituents protect brain tissue against ischemic damage in rodents.

The purpose of the present study was to test whether kava extract and its constituents kawain, dihydrokawain, methysticin, dihydromethysticin and yangonin provide protection against ischemic brain damage. To this end, we used a model of focal cerebral ischemia in mice and rats. Ischemia was induced by microbipolar coagulation of the left middle cerebral artery (MCA). To quantify the size of the lesion in mice, the area of the infarct on the brain surface was assessed planimetrically 48 h after MCA occlusion by transcardial perfusion of carbon black. In the rat model infarct volume was determined 48 h after MCA occlusion by planimetric analysis and subsequent integration of the infarct areas on serial coronal slices. Compounds were administered i.p., except the kava extract, which was administered orally. The effects of the kava extract and its constituents were compared with those produced by the typical anticonvulsant, memantine. The kava extract, methysticin and dihydromethysticin produced effects similar to those of the reference substance memantine. The kava extract (150 mg/kg, 1 h before ischemia) diminished the infarct area (P less than 0.05) in mouse brains and the infarct volume (P less than 0.05) in rat brains. Methysticin, dihydromethysticin (both 10 and 30 mg/kg, 15 min before ischemia) and memantine (20 mg/kg, 30 min before ischemia) significantly reduced the infarct area in mouse brains. All other compounds failed to produce a beneficial effect on the infarct area in mouse brains. In conclusion, the kava extract exhibited neuroprotective activity, which was probably mediated by its constituents methysticin and dihydromethysticin.

Animals↗
PubMed2006

Breast masses: computer-aided diagnosis with serial mammograms.

PURPOSE: To retrospectively evaluate effects of computer-aided diagnosis (CAD) involving an interval change classifier (which uses interval change information extracted from prior and current mammograms and estimates a malignancy rating) on radiologists' accuracy in characterizing masses on two-view serial mammograms as malignant or benign. MATERIALS AND METHODS: The data collection protocol had institutional review board approval. Patient informed consent was waived for this HIPAA-compliant retrospective study. Ninety temporal pairs of two-view serial mammograms (depicting 47 malignant and 43 benign biopsy-proved masses) were obtained from 68 patient files and were digitized. Biopsy was the reference standard. Eight Mammography Quality Standards Act of 1992-accredited radiologists and two breast imaging fellows assessed digitized two-view temporal pairs (in preselected regions of interest only) by estimating likelihood of malignancy and Breast Imaging Reporting and Data System (BI-RADS) category without and with CAD. Observers' rating data were analyzed with Dorfman-Berbaum-Metz (DBM) multireader multicase method. Statistical significance of differences was estimated with the DBM method and Student two-tailed paired t test. RESULTS: Average area under the receiver operating characteristic curve for likelihood of malignancy across the 10 observers was 0.83 (range, 0.74-0.88) without CAD and improved to 0.87 (range, 0.80-0.92) with CAD (P < .05). The average partial area index above a sensitivity of 0.90 for likelihood of malignancy was 0.35 (range, 0.13-0.54) without CAD and 0.49 (range, 0.18-0.73) with CAD--a nonsignificant improvement (P = .11). For BI-RADS assessment, it was estimated that with CAD, six radiologists would correctly recommend additional biopsies for malignant masses (range, 4.3%-10.6%) and five would correctly recommend reduction of biopsy (ie, fewer biopsies) for benign masses (range, 2.3%-9.3%). However, five radiologists would incorrectly recommend additional biopsy for benign masses (range, 2.3%-14.0%), and one would incorrectly recommend reduction of biopsy (4.3%). CONCLUSION: CAD involving interval change analysis of preselected regions of interest can significantly improve radiologists' accuracy in classifying masses on digitized screen-film mammograms as malignant or benign.

Adult↗
PubMed1987

Skeletal muscle blood flow in vivo: detection with rubidium-82 and effects of glucose, insulin, and exercise.

In order to assess the effects of glucose, insulin, and exercise on skeletal muscle blood flow in vivo, we measured positron emission from the thigh muscle of anesthetized rabbits after simultaneous aortic bolus injection of 82Rb and radiolabeled microspheres (15 micron diameter). Estimates of flow with 82Rb were based on first-pass regional extraction of 82Rb by skeletal muscle. Flow estimates were made serially as a function of variations in plasma glucose and insulin and changing the muscle contractile state by electrical stimulation. Flow ranged from 3.1 ml/min/100 g at rest to 71 ml/min/100 g during stimulation. There was good agreement between the two methods of flow measurement over the entire range of flows (r = 0.96 at a slope of 0.90). Flow measured by either method did not vary significantly from baseline over a range of plasma glucose from 5 to 30 mM and plasma insulin from 0 to 20 microU/ml. When flow was increased up to 20-fold by electrical stimulation there was a decrease in extraction of 82Rb proportional to the increase in flow. However, at pharmacologic levels of insulin (greater than 150 microU/ml) flow was increased twofold as measured by radiolabeled microspheres, but not as measured by rubidium. There was no apparent decrease in extraction of 82Rb with high insulin. The discrepancy between the microsphere measured flow and rubidium measured flow with high plasma insulin levels can be explained by the assumption that the expected decrease in the extraction fraction was counteracted by an increase in Na+/K+-ATPase activity. It is concluded that the first-pass flow model gives valid estimates of skeletal muscle blood flow in vivo with 82Rb, provided that plasma insulin levels are normal.

Animals↗
PubMed1998

Chromatographic and immunological identification of GnRH (gonadotropin-releasing hormone) variants. Occurrence of mammalian and a salmon-like GnRH in the forebrain of an eutherian mammal: Hydrochaeris hydrochaeris (Mammalia, Rodentia).

The molecular variants of Gonadotropin releasing hormone (GnRH) in brain extracts of the eutherian mammal Hydrochaeris hydrochaeris (Mammalia, Rodentia) were characterized. An indirect method combining reverse-phase high-performance liquid chromatography (RP-HPLC) and radioimmunoassay (RIA) with different antisera was used. Two different forebrain regions (olfactory bulbs and preoptic-hypothalamic region) were analyzed. Characterization of RP-HPLC fractions from preoptic-hypothalamic extracts with three different RIA systems revealed two immunoreactive GnRH (ir-GnRH) peaks coeluting with mammalian GnRH (mGnRH) and salmon GnRH (sGnRH) synthetic standards. These results were additionally supported by serial dilution studies with specific antisera. Similar results were obtained from olfactory bulb extracts with the same methodology. However, a third ir-GnRH peak in a similar position to that of chicken GnRH II (cIIGnRH) synthetic standard was revealed. As far as we know, this is the first report showing chromatographic and immunological evidences for the presence of a second GnRH variant in the forebrain of an eutherian mammal.

Animals↗
PubMed1977

Corneal thickness measurements following intraocular lens implantation.

Corneal thickness measurements were carried out on patients who had undergone combined phacoemulsification cataract extraction with simultaneous intraocular lens implantation in one eye at least six months prior to the date of measurement. The unoperated eye served as control. Another group of patients who had undergone simultaneous phacoemulsification cataract extraction combined with intraocular lens implantation were followed over a two-month period of time, serial corneal thickness measurements being performed. The results indicate that, in surgery performed by experienced surgeons on carefully selected patients, phacoemulsification cataract extraction using the Kelman technique combined with intraocular lens implantation had no significant effect on corneal endothelial function as reflected in corneal thickness changes.

Aged↗
PubMedDate not supplied

Treatment of chronic otitis externa with Trichophyton, Oidiomycetes, Epidermophyton antigen.

Chronic otitis externa may be a sensitivity (dermatophytid) reaction from a distant pathogenic superficial fungal infection, eg, tinea pedis and monilial vaginitis, which are often difficult to eradicate. Skin testing is done with a combined antigenic extract of TOE (Trichophyton; Oidiomycetes, ie, Monilia or Candida albicans; and Epidermophyton). The serial dilution titration method of skin testing is done, and the end point of reaction is determined. Treatment is begun on a weekly basis, with subcutaneous administration of 0.3 cc of a TOE extract two dilutions weaker than the end point. Symptomatic improvement is usually noted within two to four weeks, with objective improvement within eight weeks. After a variable maintenance period, the injections are spaced at less frequent intervals.

Antigens, Fungal↗