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5-Azacytidine-induced undercondensations in human chromosomes.

The cytosine analogue 5-azacytidine induces very distinct undercondensations in human chromosomes if applied to lymphocyte cultures. The number of induced undercondensations and their chromosomal localization can be varied by the 5-azacytidine dose and the treatment time. "Pulverized" chromosomes or undercondensations in the G-band-positive chromosome regions are produced with high doses and long treatment times. If applied in low doses during the last hours of culture, 5-azacytidine induces specific undercondensations in the heterochromatin of chromosomes 1, 9, 15, 16, and Y. Optimum conditions required for inducing the various types of undercondensation in the chromosomes were determined. Various examples of the use of 5-azacytidine in the analysis of chromosome rearrangements involving heterochromatic regions are presented.

Azacitidine↗

The human leulocyte test system. VII. Further investigations concerning micronucleus-derived premature chromosome condensation.

Premature chromosome condensation (PCC) from X-ray induced micronuclei shows a dose-effect relationship in human leukocytes in vitro. Preparations at different culture times without colcemide treatment reveal complex variations of the frequencies of micronuclei and PCC correlated with the fixation time. The positions of PCC patches in the metaphase plate and the frequencies of different PCC types (S and G2) ar independent on the X-ray dose. The latter indicates that the slowing down of the micronuclei in the cell cycle, which is the reason for the formation of PCC, may be an outcome rather of a regulatory phenomenon than of an unspecific physiological damage of the chromatin included in the micronuclei. This is especially evident from labeling experiments with tritiated thymidine, showing that the extent of asynchrony between main nuclei and micronuclei is independent on the X-ray dose. Labeling experiments with tritiated uridine reveal a X-ray dose dependent suppression of RNA synthesis in cells with main nuclei and micronuclei. THE S-phase nature of "pulverized" PCC patches could be verified by incorporation of tritiated thymidine in aound 50%. Staining of centromeric heterochromatin in micronuclei reveal a frequency of micronuclei with centromeric heterochromatin resembling the frequency of G2-phase PCC found in mitoses.

Cell Nucleus↗

Centromeric association and non-random distribution of centromeres in human tumour cells.

Centromere arrangement in interphase and metaphase cells of two human tumour cell lines was analysed using anti-kinetochore antibodies as immunofluorescent probes. In GLC1 interphase nuclei, kinetochores were non-randomly positioned around the nucleolus and close to the nuclear membrane. During S and early G2 phase, necklace-like strands of kinetochores were formed in the centre of the nucleus. The duplication of sister kinetochores during the G2 phase was not synchronized. At late G2 phase, a relatively random topological distribution of centromeres was observed with short linear arrays of sister kinetochores. Carefully spread metaphase plates of MDA-MB231 cells generally exhibited a linear alignment of centromeres and large centromeric clusters. In completely pulverized MDA-MB231 cells, centromeres showed a strong tendency to associate with each other.

Breast Neoplasms↗

Segmentation of human chromosomes induced by 5-ACR (5-azacytidine).

The 5-ACR (5-azacytidine) introduced in human lymphocyte cultures induces a lack or a delay of condensation of some chromosome segments corresponding to the G-bands. The resulting R-banding is very similar to that obtained with a 7-h treatment by BrdU, although the segmentation may be much stronger (pulverization) with high doses. However, the 5-ACR does not induce chromatid asymmetry, as BrdU does. This constitutes a new argument for considering that the segmentation and the asymmetry of chromatids depend, at least partly, on two different mechanisms, where proteins are probably involved. Another effect of 5-ACR is to increase chromosome associations by satellites, secondary constrictions, and telomeric regions.

Azacitidine↗

Changes in the solubility of enamel mineral at various stages of porcine amelogenesis.

The solubility of enamel mineral (a carbonated apatite) formed at various stages of porcine amelogenesis was investigated at controlled partial pressures of CO2. Enamel samples were obtained from the outer (young) secretory, inner (old) secretory, early (soft) and late (hard) mature enamel of the permanent dentition of slaughtered piglets. The dissected enamel was pulverized and subjected to a plasma ashing at low temperature to remove organic matter. The composition (Ca, total P, HPO4, and CO3) of the enamel mineral was determined chemically. The enamel mineral contained significant amounts of carbonate and acid phosphate; the model adopted for its stoichiometry was [Ca]5-x [HPO4]v[CO3]w[PO4]3-x[OH]1-x. Each enamel sample was equilibrated in dilute phosphoric acid solutions (0.01-1.2 mM) under Pco2 = 1.86 and 1.75%. Equilibration of the enamel samples usually took 20-25 days; the solution composition (pH, concentrations of Ca, P, Mg, Na, and K, and activity of Ca2+) was determined periodically. The composition of the solution at equilibrium showed that (1) the outer (younger) secretory mineral was the most soluble and the solubility of enamel mineral decreased with advancing developmental stages; (2) the mean activity product in the saturated solutions for the outer secretory enamel was the same as that calculated on the basis of the reported composition of the enamel fluid; and (3) the solubility data obtained with most of the enamel samples were consistent with a model in which the equilibration includes two processes: dissolution of the original enamel mineral and precipitation of a new carbonatoapatite. Analyses of the equilibrated samples, particularly the mature enamel, by electron microscopy, supported the precipitation of carbonatoapatite.(ABSTRACT TRUNCATED AT 250 WORDS)

Amelogenesis↗

Competitive adsorption of magnesium and calcium ions onto synthetic and biological apatites.

Magnesium (Mg) is a conspicuous constituent of hard tissues but its possible role in biomineralization is poorly understood. It is possible that Mg2+ adsorbed onto bioapatites may contribute to the modulation of crystal growth as such inhibitory activity has been reported for synthetic apatites. The present study was undertaken to determine the adsorption isotherms of Mg ions onto synthetic apatites and biominerals in tooth and bone tissues in the presence of other ions of natural occurrence. Synthetic crystals used as adsorbents were hydroxyapatite and, as a better prototype for the biomineral, Mg-containing carbonatoapatite. Human enamel and dentin materials were obtained from extracted, caries-free, permanent teeth. Porcine dentin materials at two developmental stages were obtained from erupted deciduous and unerupted permanent teeth of a 6-month-old slaughtered piglet. Porcine bone was obtained from the cortical portion of the mandible of the same animal. All biomineral samples were pulverized and then treated by plasma ashing (deproteination) at about 60 degrees C. Each of the powdered samples was equilibrated in solutions containing various initial concentrations of Mg2+, Ca2+, and Na+ (or K+) as nitrate salts. Following equilibration, concentrations (and activities) of magnesium and calcium ions in the experimental solution were determined. The pH values of the equilibrium solutions were in the range of 6.2-6.5. Experimental data of the Mg adsorption onto hydroxyapatite were interpreted on the basis of a Langmuir-type model for binary systems assuming competition of Mg2+ and Ca2+ for the same adsorption sites on the crystal surfaces of the apatites.(ABSTRACT TRUNCATED AT 250 WORDS)

Adsorption↗

The effects of caffeine on the maxillary composition in the newborn rat.

The possible influence of caffeine on maxillary structure was studied. Seventeen pregnant rats at days 9 of gestation were randomly divided into two groups. The dams of group 1 received a 20% protein diet ad libitum throughout the experimental period. The dams of group 2 were pair-fed, with group 1, a 20% protein diet supplemented with 2 mg/100 g body weight (B.W.) caffeine. At birth, pups were mixed within the same group and 8 randomly selected pups were assigned to each dam and continuously fed the respective diet. On day 22, 11 male pups from the control and 12 males from the caffeine group were randomly selected, separated from the dams, and continued to be fed their respective diets. On day 44, a rubber elastic band was inserted between the first and second maxillary right molars. The size of the elastic band was increased throughout the next 5 days. Animals were sacrificed at day 49 and the composition of the maxillas was analyzed. After pulverization, organic and inorganic contents of the bones were measured. Zinc (Zn) and hydroxyproline concentration of the caffeine group showed a significant decrease when compared with those of the controls. However, Ca, P, Mg, and hexosamine concentration showed no difference between the groups. The interdental space measured occlusally and laterally with the visual method, and occlusally in histological sections showed no significant difference between the control and caffeine groups, although variation of the space in the caffeine group was less than in the control group.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Glycosaminoglycans in the human aorta in diabetes mellitus: a study of tunica media from areas with and without atherosclerotic plaque.

Alterations in the connective tissue of the arterial wall have been suggested to play a role in the development of macrovascular disease in diabetes mellitus. The present study deals with changes in the content of GAG in aortic tunica media in human diabetes by separately analysing normal areas and areas with fibrous plaques. The thoracic aorta from 15 diabetic patients (7 with IDDM, 8 with NIDDM), and 30 sex- and age-matched non-diabetic subjects were collected at autopsy. Tunica intima was removed and GAG were isolated from the dried defatted and pulverized tunica media. GAG were quantified by uronic acid analysis and characterized by electrophoresis on cellulose acetate. Results showed that IDDM patients had a relative and absolute increase in hyaluronic acid in normal tunica media compared to non-diabetic subjects. There was a significant positive correlation between hyaluronic acid content of normal tunica media and duration of diabetes, but not between hyaluronic acid content and age. When tunica media from plaque areas was compared to normal areas the same pattern was evident in diabetic patients as in non-diabetic patients--significantly increased proportion of dermatan sulphate and reduced hyaluronic acid. The data agree with the notion that the arterial wall is subject to different pathological processes in diabetes, one of classical atherosclerosis with changes in GAG similar to non-diabetic subjects, and the other seen in areas without plaques with dissimilar alterations in GAG. These data therefore support the concept of the presence of a macrovascular disease in diabetes different from atherosclerosis.

Aged↗

Aortic collagen alterations in human diabetes mellitus. Changes in basement membrane collagen content and in the susceptibility of total collagen to cyanogen bromide solubilisation.

Structural alterations in arterial extracellular matrix components have been suggested to play a role in the development of arterial disease among patients with diabetes mellitus. This study examines the quantity and quality of collagenous components in aortas from diabetic patients. In order to obtain data about the arterial tissue concentration of type IV and V collagen in diabetic and non-diabetic patients, aortas from 21 patients with diabetes (9 with Type 1 (insulin-dependent) diabetes and 12 with Type 2 (non-insulin-dependent) diabetes), were collected at autopsy together with aortas from groups of sex- and age-matched patients. Intima and media samples from normal and fibrous plaque areas from the individual vessels were evaluated. Pulverized, dried and defatted tissue samples were subjected to chemical solubilization with cyanogen bromide and subsequent immuno-chemical quantitation of the dissolved type IV and V collagen in an ELISA. It was found that the concentration of type IV collagen was increased in the tunica media both in plaque and non-plaque areas in the samples from the diabetic patient groups as compared to the non-diabetic groups. No consistent differences in type IV collagen concentrations were found between diabetic and non-diabetic patients in tunica intima. The type V collagen concentrations and the total collagen content were not altered in the diabetic samples. The fraction of the total collagen that was solubilized during cyanogen bromide treatment was determined, and it was found that this fraction was decreased in most tissue areas in the diabetic patient groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

Mastication with and without removable partial dentures: an intraindividual study.

The chewing of 22 subjects, with a total of 27 removable partial dentures (RPD) that had been used for more than 6 months, was analyzed by four different methods. The study design was an intraindividual comparison of chewing, with and without the RPDs. Chewing tests were a color mixture of two-colored gums, chewing gum bolus shaping, comminution of Optosil silicone tablets, and number of chewing strokes to first swallow of an almond. The participants were also asked some standardized questions about their eating, biting, and chewing capacity. There was no significant difference in efficiency as measured by chewing gum color mixing, chewing gum shape, and pulverization of Optosil between chewing with and without the RPDs. Significantly more strokes were needed to swallow an almond without than with the RPD in place. With their RPDs in place, all subjects could chew a sandwich and an apple, and all but one could chew whole meat. The study shows that the chewing process should preferably not be analyzed by one method only.

Adult↗

Steatorrhea, a simple and rapid method of diagnosis.

A simple and rapid method for the diagnosis of steatorrhea is described. The method is based on the assessment of the difference of specific gravity and lipophilic property between a normal stool and that in steatorrhea, using a benzene-NaBr system. The stool samples are dried, pulverized, and then investigated. Steatorrhea is defined either as a fat or more in a 24-hour sample. Stool samples of 71 infants and children were examined. Among these were 23 healthy subjects, 10 with upper respiratory tract infections, and 6 with pneumonia. A further 31 children had celiac disease, 9 of them were on a gluten-free diet, and one child had cystic fibrosis. The procedure described renders single stool samples suitable for the assessment of steatorrhea. As a screening test the method proved to be rapid and accurate in the diagnosis of steatorrhea.

Adolescent↗

Cardioprotective actions of wild garlic (allium ursinum) in ischemia and reperfusion.

The susceptibility to ventricular arrhythmias under the conditions of cardiac ischemia and reperfusion was investigated in the Langendorff heart preparation of rats fed for eight weeks a standard chow enriched with 2% of pulverized wild garlic leaves. The isolated hearts were perfused with a modified Krebs-Henseleit solution. The incidence of ventricular fibrillation (VF) during 20 min occlusion of the descending branch of the left coronary artery (LAD) was significantly reduced in the wild garlic group as compared to untreated controls (20% vs 88%). The same holds for the size of the ischemic zone (33.6% vs 40.9% of heart weight). In the reperfusion experiments (5 min after 10 min ischemia), ventricular tachycardia (VT) occurred in 70% of the wild garlic group vs 100% in untreated controls and VF in 50% vs 90%. The time until occurrence of extrasystoles, VT or VR was prolonged. No significant alterations in cardiac fatty acid composition could be observed. Although the prostacyclin production was slightly increased in hearts of the wild garlic group, inhibition of cyclooxygenase by acetylsalicylic acid (ASA; aspirin) could not completely prevent the cardioprotective effects suggesting that the prostaglandin system does not play a decisive role in the cardioprotective action of wild garlic. Furthermore, a moderate angiotensin converting enzyme (ACE) inhibiting action of wild garlic was found in vitro as well as in vivo that could contribute to the cardioprotective and blood pressure lowering action of wild garlic. Whether a free radical scavenging activity of wild garlic is involved in its cardioprotective effects remains to be established.

Angiotensin-Converting Enzyme Inhibitors↗

Isolation of bovine intramuscular connective tissue.

This report presents a simple and rapid method for separating intramuscular connective tissue (IMCT) from bovine muscles. After lyophilization, muscle samples are blended and fibrous connective tissue is isolated from pulverized muscle protein by sieving. This method requires little expense in time and equipment, is easy to handle and therefore, is well suited to large sample output. The isolation of IMCT is necessary for further studies on intramuscular collagen. Recoveries and collagen enrichment of this method have been tested using several bovine muscles varying in collagen content.

Animals↗

Fatty acid composition and arachidonic acid concentrations in alveolar bone of rats fed diets with different lipids.

The purpose of the present study was to determine if the type of dietary fat can modify the fatty acid composition and arachidonic acid levels in the alveolar bone phospholipids. Three groups of rats were fed nutritionally adequate semipurified diets containing different lipids: 10% corn oil (control, group I, rich in n-6 fatty acids); 9% butter + 1% corn oil (experimental, group II, rich in saturated fatty acids); and 9% ethyl ester concentrate of n-3 fatty acids + 1% corn oil (experimental, group III, rich in n-3 fatty acids). After 10 weeks of feeding the various diets, rats were killed, maxillae and mandibles were dissected out, and the soft tissue was removed. Bone was frozen in liquid nitrogen and pulverized. Powdered bone was extracted for total lipids, and phospholipids were isolated by column chromatography. The fatty acid composition and arachidonic acid concentrations were determined in total phospholipids after the addition of an internal standard, octadecatetraenoic acid (18:4n-3), and subsequent gas chromatography. The type of dietary lipids had a profound influence on the fatty acid composition of bone lipids. Arachidonic acid concentrations were significantly lower in total phospholipids of mandibles and maxillae of rats fed the experimental diets than in those fed the control diet. Because arachidonic acid is a precursor of prostaglandin E2 and leukotriene C4, a significant reduction in its concentration may result in reduced levels of these eicosanoids in the alveolar bone.

Alveolar Process↗

Dietary lipids, prostaglandin E2 levels, and tooth movement in alveolar bone of rats.

A previous study showed that certain dietary lipids can alter arachidonic acid concentrations in alveolar bone. Because arachidonic acid is a precursor of prostaglandin (PG) E2, which is known to play an important role in orthodontic tooth movement, the purpose of the present study was to determine the effect of dietary lipids on PGE2 levels and tooth movement. Two groups of male Sprague-Dawley rats (20/group) were fed nutritionally adequate purified diets containing 10% corn oil (group I, rich in n-6 fatty acids) or 9% ethyl ester concentrate of n-3 fatty acids + 1% corn oil (group II rich in n-3 fatty acids). After 5 weeks of feeding the diets, orthodontic force of 56 g was applied to the maxillary incisors to tip them distally. Prior to killing the rats at day 4 and 8 of orthodontic force application, tooth movement was measured by computerized image analysis. Premaxillae were dissected out free of soft tissue and incisors. The alveolar bone was frozen in liquid nitrogen, pulverized, and lipids were extracted. The concentrations of arachidonic acid and fatty acid composition of total phospholipids were measured by gas chromatography. PGE2 levels were measured by enzyme immunoassay. Arachidonic acid and PGE2 concentration were significantly lower (P < 0.001) in alveolar bone of rats in group II than in group I. The tooth movement was also significantly lower (P < 0.02) in group II than in group I at both 4 and 8 days. The results suggest that PGE2 levels in alveolar bone and orthodontic tooth movement can be affected by the type of dietary fat.

Alveolar Process↗

Haematogenous Serratia marcescens endophthalmitis in an HIV-infected intravenous drug addict.

A case of haematogenous Serratia marcescens endophthalmitis in an HIV-infected intravenous drug addict is described. The patient was admitted with fever, ocular pain and visual loss in the right eye following an i.v. injection of pulverized buprenorphine. A vitreous humor culture grew S. marcescens. The patient was treated with i.v. ceftriaxone (2 g b. i. d.), i.v. amikacin (500 mg b. i. d.) and p. o. fosfomycin (1 g q. i. d.) for three weeks. The ocular infection was cured, although the visual function was lost, leading to blindness. To our knowledge, this is the second case in the reviewed Anglo Saxon literature of S. marcescens endophthalmitis in parenteral drug addicts.

Adult↗

Intravascular ultrasound imaging for guidance of atherectomy and other plaque removal techniques.

Intravascular ultrasound imaging provides a direct view of atherosclerotic disease, generating in vivo information about the depth and mechanical characteristics of plaque at any point in the vessel wall. For this reason, ultrasound has significant potential to serve as a guidance modality for catheter-based techniques designed to remove or ablate plaque. Although the current generation mechanical atherectomy, laser ablation and ultrasound pulverization techniques all have some specificity for attacking plaque as opposed to normal vessel wall, it appears that in practice all of these devices will continue to carry a risk of traumatizing or even perforating arteries. In addition, it seems highly likely that aggressive 'debulking' of plaque will require some type of guidance beyond angiography - a role which ultrasound is theoretically well suited to play. The purpose of this review is to consider the theoretical and practical applications of ultrasound imaging as a guide to catheter-based plaque removal and ablation techniques. Specific uses will be discussed with respect to both directional and coaxial therapeutic devices.

Arteriosclerosis↗

Cytogenetic toxicity of D2O in human lymphocyte cultures. Increased sensitivity in Fanconi's anemia.

Chromosomal aberrations were scored in lymphocyte cultures from healthy individuals, patients with Bloom syndrome, and patients with Fanconi's anemia, after 4-5 h exposure to culture medium containing 90% heavy water (D2O). D2O treatment resulted in occasional pulverization of metaphases, and increased frequencies of chromosomal breakage. Patients with Fanconi's anemia were particularly sensitive to the chromosome breaking effect of D2O.

Anemia, Aplastic↗