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Macronuclei and micronuclei in Tetrahymena thermophila contain high-mobility-group-like chromosomal proteins containing a highly conserved eleven-amino-acid putative DNA-binding sequence.

HMG (high-mobility-group protein) B and HMG C are abundant nonhistone chromosomal proteins isolated from Tetrahymena thermophila macronuclei with solubilities, molecular weights, and amino acid compositions like those of vertebrate HMG proteins. Genomic clones encoding each of these proteins have been sequenced. Both are single-copy genes that encode single polyadenylated messages whose amounts are 10 to 15 times greater in growing cells than in starved, nongrowing cells. The derived amino acid sequences of HMG B and HMG C contain a highly conserved sequence, the HMG 1 box, found in vertebrate HMGs 1 and 2, and we speculate that this sequence may represent a novel, previously unrecognized DNA-binding motif in this class of chromosomal proteins. Like HMGs 1 and 2, HMGs B and C contain a high percentage of aromatic amino acids. However, the Tetrahymena HMGs are small, are associated with nucleosome core particles, and can be specifically extracted from macronuclei by elutive intercalation, properties associated with vertebrate HMGs 14 and 17, not HMGs 1 and 2. Thus, it appears that these Tetrahymena proteins have features in common with both of the major subgroups of higher eucaryotic HMG proteins. Surprisingly, a linker histone found exclusively in transcriptionally inactive micronuclei also has several HMG-like characteristics, including the ability to be specifically extracted from nuclei by elutive intercalation and the presence of the HMG 1 box. This finding suggests that at least in T. thermophila, proteins with HMG-like properties are not restricted to regions of transcriptionally active chromatin.

Amino Acid Sequence↗

Mutual synaptic associations between neurons containing neuropeptide Y and neurons containing enkephalin in the arcuate nucleus of the rat hypothalamus.

We determined reciprocal synaptic associations between neurons containing neuropeptide Y (NPY) and neurons containing enkephalin-8 peptide (Enk-8) in the arcuate nucleus (ARC) of the rat hypothalamus by using a double-staining immunocytochemical method. Animals treated with an intraventricular administration of colchicine and animals treated with a hemicomplete cut of the medial basal hypothalamus were used. We principally labeled NPY with silver-gold particles and Enk-8 with diaminobenzidine (DAB) chromogen but in part labeled reversely the two antigens to obtain better visualization of Enk-8 cell bodies. Neurons immunoreactive for NPY were found in the mediobasal area, while those for Enk-8 were located somewhat laterally. Their fibers, however, were densely distributed throughout the ARC in a similar fashion. Electron microscopically, we found synaptic junctions between axonal terminals of Enk-8 neurons and perikarya of NPY neurons and between axonal terminals of NPY neurons and perikarya of Enk-8 neurons. In addition, we found synaptic junctions between immunoreactive Enk-8 cell bodies and processes and also between immunoreactive NPY cell bodies and processes. These interneuronal associations may contribute to the interaction of information coming in or going out from this nucleus.

Animals↗

Comparative three-year caries protection from an aluminum-containing and a fluoride-containing toothpaste.

In the present study, 2,087 fifth-graders (mean age: 12 years) from two cohorts entered a 3-year clinical toothpaste trial. The study subjects were randomly allocated to two groups: one group was a positive control using a 0.83% NaFPO3 toothpaste, while the other group brushed with a test toothpaste containing 1.77% KAl(SO4). Both toothpastes were used unsupervised at home. One set of bite-wing radiographs was obtained at the beginning of the trial and another at the 3-year follow-up. Clinical registrations of caries were performed by the municipal dentists responsible for the child's dental care. After 3 years, the mean caries increment was significantly higher in the group using the aluminum-containing toothpaste measured both clinically and radiographically.

Alum Compounds↗

Expression of apoptosis-associated speck-like protein containing a caspase recruitment domain, a pyrin N-terminal homology domain-containing protein, in normal human tissues.

Apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) is a pyrin N-terminal homology domain (PYD)- and caspase recruitment domain (CARD)-containing a proapoptotic molecule. This molecule has also been identified as a target of methylation-induced silencing (TMS)-1. We cloned the ASC cDNA by immunoscreening using an anti-ASC monoclonal antibody. In this study, we determined the binding site of the anti-ASC monoclonal antibody on ASC and analyzed the expression of ASC in normal human tissues. ASC expression was observed in anterior horn cells of the spinal cord, trophoblasts of the placental villi, tubule epithelium of the kidney, seminiferous tubules and Leydig cells of the testis, hepatocytes and interlobular bile ducts of the liver, squamous epithelial cells of the tonsil and skin, hair follicle, sebaceous and eccrine glands of the skin, and peripheral blood leukocytes. In the colon, ASC was detected in mature epithelial cells facing the luminal side rather than immature cells located deeper in the crypts. These observations indicate that high levels of ASC are abundantly expressed in epithelial cells and leukocytes, which are involved in host defense against external pathogens and in well-differentiated cells, the proliferation of which is regulated.

Animals↗

GDNF increases the density of cells containing calbindin but not of cells containing calretinin in cultured rat and human fetal nigral tissue.

Among the dopaminergic neurons in substantia nigra pars compacta and in the ventral tegmental area, subpopulations express the calcium-binding proteins calbindin (CB) and calretinin (CR), and the CB-containing neurons are supposed to be less prone to degeneration in Parkinson's disease. Glial cell line-derived neurotrophic factor (GDNF) is a potent survival factor for nigrostriatal dopaminergic neurons. Using free-floating roller-tube (FFRT) cultures derived from fetal rat (E14) ventral mesencephalon we found that GDNF (10 ng/ml) significantly increased the number of surviving tyrosine hydroxylase (TH)-immunoreactive neurons. The possible effects of GDNF treatment on CB-immunoreactive (CB-ir) and CR-ir neurons in such cultures were examined in the present study. The neuronal cell densities were measured by quantifying the numbers of CB-ir and CR-ir neurons in areas of sections through the most extensive parts of the spherical cultures. In 4-day-old and 8-day-old cultures GDNF treatment increased the density of CB-ir neurons by 50% and 59%, respectively. Partial co-existence of TH and CB was shown using the method of double immunolabeling. The density of CR-containing neurons was unaffected by GDNF treatment as confirmed by Western blotting for CR. Parallel effects of GDNF treatment were obtained for cultures of human fetal ventral mesencephalon (8 weeks postconception). In conclusion, our findings identify GDNF as a potent factor for fetal rat and human nigral CB-ir neurons able to promote their survival in culture. Referring to a suggested neuroprotective role of CB, the results may be of relevance in the context of neuronal transplantation of patients suffering from severe Parkinson's disease.

Animals↗

Metal-containing components in medicinal plants. III. Manganese-containing components in Theae folium as oral magnetic resonance imaging contrast materials.

A manganese-containing component from the water-extract of Theae folium (green tea) was found to exert an augmentative effect in magnetic resonance imaging (MRI) contrast and may possibly be a manganese(II) complex with a pectin-like polysaccharide capable of shortening the spin-lattice relaxation time (T1) of water protons. Even though only the manganese(II) ion with S = 5/2 is active in T1-shortening ability, which should enhance contrast, complexation of this ion with the polysaccharide causes a marked increase in its activity. This manganese-containing pectin-like polysaccharides should prove useful as a low-toxic oral gastrointestinal contrast material in MRI.

Chromatography, High Pressure Liquid↗

Fluorescent phospholipid analogs as microscopic probes for detection of the mycolic acid-containing layer in Corynebacterium glutamicum: detecting alterations in the mycolic acid-containing layer following ethambutol treatment.

Corynebacterium glutamicum belongs to the mycolic acid-containing actinomycetes, which also include Mycobacterium, Nocardia, and Rhodococcus. The cells of this group possess a cell wall with a thick outer layer composed primarily of mycolic acid, which functions as a permeability barrier. To investigate the mechanism of mycolic acid-containing layer (mycolate layer) formation, we have developed a fluorescence microscopic technique detecting the mycolate layer in situ. The staining specificity of fluorescence-labeled phospholipid analogs was determined by simultaneous staining with the hydrophobic fluorescent dye Nile Red and peptidoglycan-staining fluorescence-conjugated vancomycin. We found that fluorescence-labeled phospholipid analogs preferentially stain the mycolate layer. Using this technique, we observed the effect of the anti-mycobacterial drug ethambutol on C. glutamicum mycolate-layer formation. Ethambutol interfered specifically with mycolate-layer formation on the division planes and cell poles, while the side-wall mycolate layer was not severely affected. This indicates that mycolate-layer formation occurs mainly on division planes and cell poles in C. glutamicum, where the peptidoglycan layer is actively synthesized.

Anti-Infective Agents↗

Genetic analysis of the Caenorhabditis elegans pax-6 locus: roles of paired domain-containing and nonpaired domain-containing isoforms.

PAX-6 proteins are involved in eye and brain development in many animals. In the nematode Caenorhabditis elegans the pax-6 locus encodes multiple PAX-6 isoforms both with and without a paired domain. Mutations in the C. elegans pax-6 locus can be grouped into three classes. Mutations that affect paired domain-containing isoforms cause defects in epidermal morphogenesis, epidermal cell fates, and gonad cell migration and define the class I (vab-3) complementation group. The class II mutation mab-18(bx23) affects nonpaired domain-containing isoforms and transforms the fate of a sensory organ in the male tail. Class III mutations affect both paired domain and nonpaired domain isoforms; the most severe class III mutations are candidate null mutations in pax-6. Class III mutant phenotypes do not resemble a simple sum of class I and class II phenotypes. A comparison of class I and class III phenotypes indicates that PAX-6 isoforms can interact additively, synergistically, or antagonistically, depending on the cellular context.

Animals↗

Copper-zinc containing and manganese containing superoxide dismutase in the ground squirrel/Citellus citellus/--the effect of hibernation.

The distribution of Copper-Zinc containing and Manganese-containing superoxide dismutase in the liver, kidney, interscapular brown adipose tissue (IBAT) and brain of the ground squirrel, as well as the effect of hibernation, was studied. Activity of both forms of SOD was highest in the liver and lowest in the brain. Activity of the Mn SOD in relation to total SOD was higher in the liver and kidney of the ground squirrel as compared with results reported for other rodents. The highest activity of Mn SOD in relation to total SOD was found in the IBAT and brain (36% and 49%, respectively). Total SOD activity per mg proteins and per g wet mass in IBAT and brain of hibernating animals was increased: for IBAT, p less than 0.05 and p less than 0.025, respectively; for brain, p less than 0.01 and p less than 0.025, respectively. Protein content in hibernating ground squirrel was not significantly changed. In the hibernating ground squirrel CuZn SOD activity in IBAT and brain was higher as compared with the active animal (p less than 0.025 and p less than 0.005, respectively). In the liver and kidney CuZn SOD was not significantly changed during the hibernation. In the liver and brain of the hibernating animals a lower Mn SOD activity was found (p less than 0.005 and p less than 0.05, respectively).

Adipose Tissue, Brown↗

Activity of immunoglobulin G-coated red cell ghosts containing pentamidine against macrophage-contained Leishmania in vitro.

Pentamidine is an antileishmanial agent that is often toxic at therapeutic dosages. The obligate intramacrophage localization of Leishmania indicates that encapsulation of pentamidine within a carrier phagocytized by macrophages (IgG-coated sheep red cell ghosts) might improve activity. In in vitro experiments, treatment of infected mouse macrophages for 1 hour with a mean of 1.4 micrograms of encapsulated drug resulted in a calculated drug concentration of 180 micrograms/ml macrophage, and in 73% suppression of organism multiplication within the macrophages after 4-5 days of further cultivation. In comparison, 27 micrograms unencapsulated drug/ml was needed for similar suppression. Electron microscopic examination 5 hours after phagocytosis of IgG-ghosts revealed that 95% of organisms were adjacent to ghosts in phagolysosomes. Fusion of drug carrier with phagolysosome containing drug target is therefore an important step in carrier-mediated parasite suppression in this model. These results suggest that IgG-coated erythrocyte ghosts containing pentamidine have potential as an antileishmanial formation.

Amidines↗

Small HDL particles containing two apoA-I molecules are precursors in vivo to medium and large HDL particles containing three and four apoA-I molecules in nonhuman primates.

We hypothesized that small HDL particles, containing two apoA-I molecules but no apoA-II (LpAI), may be converted in vivo into medium and large HDL particles, containing three or four apoA-I molecules, respectively, and that more conversion will occur in animals with higher HDL concentrations. To test this possibility, kinetic studies of small LpAI were performed in African green monkeys with either high plasma HDL cholesterol concentrations (120 +/- 36 mg/dl, mean +/- SD, n = 3) or low plasma HDL cholesterol concentrations (40 +/- 13 mg/dl, n = 3). Tracer small LpAI was purified, without ultracentrifugation, by immunoaffinity and gel filtration. After injection, the specific activity of apoA-I in small, medium, and large HDL, consisting of both LpAI and LpAI:AII particles, was followed. A multicompartmental model was developed with the simultaneous analysis of urine and plasma turnover data for the kinetics of apoA-I in small, medium, and large HDL. These analyses indicated that small HDL is converted to either medium or large HDL with little or no interconversion of medium HDL and large HDL. Much of the metabolic conversion of small HDL occurs in a sequestered pool, effectively outside the circulating plasma, in a unidirectional manner before reentering the circulating plasma as medium or large HDL. The mean fractional catabolic rate of apoA-I in small, medium, and large HDL was not different comparing the high and low HDL group. In contrast, the mean production rate of apoA-I was greater in the high HDL group compared with the low HDL group. These data support the hypothesis that the plasma concentration of HDL is primarily a function of the rate of appearance of apoA-I in medium and large HDL.

Animals↗

The comparative anticaries efficacy of a dentifrice containing 0.3% triclosan and 2.0% copolymer in a 0.243% sodium fluoride/silica base and a dentifrice containing 0.243% sodium fluoride/silica base: a two-year coronal caries clinical trial on adults in Israel.

The purpose of this two-year clinical study was to provide a comparison of the anticaries efficacy associated with two commercially available, American Dental Association-accepted dentifrices: Colgate Total Toothpaste, which contains 0.3% triclosan and 2% copolymer in a 0.243% sodium fluoride/silica base; and Crest Cavity Fighting Toothpaste with Fluoristat, which contains 0.243% sodium fluoride in a silica base. The study was conducted in harmony with the published 1988 American Dental Association guidelines for studies geared toward the comparison of fluoride dentifrices. The study employed a double-blind parallel-group design, and involved adults living in communities throughout Israel. Qualifying subjects were randomly assigned to the two treatment groups, with multiple subjects in the same household all assigned to the dentifrice randomly allocated to the first among them. Caries examinations were conducted in accordance with U.S. Food and Drug Administration guidelines for the clinical evaluation of drugs to prevent dental caries. Two calibrated examiners performed all of the measurements. After treatment assignment, study participants were instructed to brush their teeth at home with their assigned dentifrice at least twice daily. Brushing instructions were reinforced by indoctrination in proper oral hygiene techniques by dental professionals, supplemented by pamphlets supplied by the sponsor and yearly mailings to participants, emphasizing good oral hygiene and the need to ensure compliance with the study. Post-baseline examinations were performed after one year of product use, and again after two years of product use. Three-thousand, three-hundred and ninety-two (3,392) subjects completed this two-year study. For these subjects, the mean caries scores (DFS, decayed or filled surfaces) at baseline were 21.96 for the Colgate Total Toothpaste group, and 21.49 for the Crest Cavity Fighting Toothpaste with Fluoristat group. For caries increment after one year, the respective means were 1.37 for the Colgate Total Toothpaste group, and 1.56 for the Crest Cavity Fighting Toothpaste with Fluoristat group. After two years, the mean caries increments were 1.46 for the Colgate Total Toothpaste group, and 1.75 for the Crest Cavity Fighting Toothpaste with Fluoristat group. No statically significant difference was indicated between the treatment groups at baseline. However, for both the one-year and two-year increments, there was a statistically significant difference between treatment groups. Relative to the Crest Cavity Fighting Toothpaste with Fluoristat group, the Colgate Total Toothpaste group presented a 12.2% reduction in caries increment scores at one year, and a 16.6% reduction in caries increment scores at two years. In accordance with the procedures and standards provided by the published guidelines of the American Dental Association for the comparison of the anticaries efficacy of fluoride dentifrices, the results of this study support the conclusion that Colgate Total Toothpaste provides a superior level of coronal anticaries efficacy compared to Crest Cavity Fighting Toothpaste with Fluoristat.

Adult↗

Adult rodent neurogenic regions: the ventricular subependyma contains neural stem cells, but the dentate gyrus contains restricted progenitors.

Neurogenesis persists in two adult brain regions: the ventricular subependyma and the subgranular cell layer in the hippocampal dentate gyrus (DG). Previous work in many laboratories has shown explicitly that multipotential, self-renewing stem cells in the subependyma are the source of newly generated migrating neurons that traverse the rostral migratory stream and incorporate into the olfactory bulb as interneurons. These stem cells have been specifically isolated from the subependyma, and their properties of self-renewal and multipotentiality have been demonstrated in vitro. In contrast, it is a widely held assumption that the "hippocampal" stem cells that can be isolated in vitro from adult hippocampus reside in the neurogenic subgranular layer and represent the source of new granule cell neurons, but this has never been tested directly. Primary cell isolates derived from the precise microdissection of adult rodent neurogenic regions were compared using two very different commonly used culture methods: a clonal colony-forming (neurosphere) assay and a monolayer culture system. Importantly, both of these culture methods generated the same conclusion: stem cells can be isolated from hippocampus-adjacent regions of subependyma, but the adult DG proper does not contain a population of resident neural stem cells. Indeed, although the lateral ventricle and other ventricular subependymal regions directly adjacent to the hippocampus contain neural stem cells that exhibit long-term self-renewal and multipotentiality, separate neuronal and glial progenitors with limited self-renewal capacity are present in the adult DG, suggesting that neuron-specific progenitors and not multipotential stem cells are the source of newly generated DG neurons throughout adulthood.

Aging↗

[Somatostatin-containing fibers and neurophysin-containing fibers in the median eminence of the fox, as seen with a double staining cytoimmunoenzyme technique].

In the external layer of the median eminence of the fox, the somatostatin-containing fibers and neurophysin-containing fibers of the hypothalamo-infundibular tract are located in distinct areas. In the neural lobe, somatostatin-positive areas are simultaneously neurophysin-positive. Outside the SON and PVN, some somatostatin-positive and neurophysin-negative perikarya are scattered close to the third ventricle. These facts suggest the existence of two somatostatin systems: a hypothalamo-infundibular (neurophysin-negative) one and a hypothalamo-neurohypophyseal (neurophysin-positive) one.

Animals↗

[Study on transgenic elite indica container line D297B containing foreign insect-resistant gene gna].

It was reported in this article that the transgenic elite indica container line D297B containing snowdrop lectin (Galanthus nivalis agglutinin, GNA) gene gna was obtained by biolisties. PCR, Southern blotting and Western blotting indicated that the transgenes were integrated into the genome of D297B and expressed in transgenic plants. Analysis of protein activity showed that product of transgene had activity of agglutinin. Resistance of transgenic seeds to Hygromycin B suggested that the transgenes were integrated in a single locus and inherited according to the pattern of 3:1 in most of transgenic plants. In addition, the PCR analysis revealed that the insect-resistant gene gna and selective marker gene hpt were co-integrated and co-inherited.

Animals↗

[Hyperthyreosis and hypothyreosis after use of iodine-containing natural products and iodine-containing vitamin and mineral supplements].

Excess iodine is known to affect thyroid function, and in predisposed individuals even a moderately increased iodine intake may cause hypothyreosis or hyperthyreosis. In our material, one patient developed hypothyreosis and five patients hyperthyreosis after using commercial health products containing iodine. The daily intake of iodine was 100-300 micrograms for a period of four months to four years. Except for a positive antibody titer to microsomal thyroid-antigen no obvious predisposing conditions were present. The thyroid function normalized two to three months after withdrawal of the iodine product. The hyperthyroid patients accounted for about 5% of all cases of hyperthyrosis seen in our hospitals during the period in question (one and a half years). It is concluded that "health products", containing iodine should be subjected to public control and the iodine content regulated. This would mean changing the present practice of no such regulation.

Adult↗

The sodium ion translocating oxalacetate decarboxylase of Klebsiella pneumoniae. Sequence of the biotin-containing alpha-subunit and relationship to other biotin-containing enzymes.

The gene encoding the alpha-subunit of the Na+ pump oxalacetate decarboxylase of Klebsiella pneumoniae was cloned and sequenced. The deduced primary structure of the protein was confirmed by protein sequencing of about 30% of the polypeptide chain. The gene has a GC content of 67% and codes for 596 amino acids. The N-terminal methionine is removed in the mature protein which has a calculated molecular mass of 63,600 daltons. The protein consists of two different domains that are connected by a stretch of amino acid residues susceptible to proteolytic cleavage. Limited proteolysis of the native enzyme with trypsin produced fragments of about 51 kDa and 10.2 kDa, the latter of which started with valine 491 and contained the biotin prosthetic group. Peptide sequencing indicated binding of the biotin prosthetic group to lysine 561, 35 residues from the C terminus. The decarboxylase contains an extended alanine- and proline-rich region (positions 502-532) on the N-terminal side of the 10.2-kDa biotin domain. This sequence includes a total of 16 alanine and 9 proline residues.

Alanine↗

[The effect of visible light on the stability of isoprenaline, epinephrine and levarterenol solutions in various containers. 86. Problems in the use of plastic containers for liquid preparations. 21. The stability of drugs and preparations].

The stability of solutions of phenylalkylamines of the benzcatechine type was investigated in containers of different structure under influence of diffuse day light, direct sun light and artificial visible light. Thereby a combined influence of different the decomposition increasing factors could be observed. The decompositions were slowest in containers of PE-ND with pigment additionals and were increased from PE-HD, PE-ND, colourless glas up to brown glas .

Drug Packaging↗