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At least 955 records · Page 53Linked to original sources
PubMed1987

Properties of L-streptococci in comparison with those of A-streptococci.

Despite some similarities, L-streptococci could be clearly differentiated from A-streptococci. Formamide, autoclaved and nitrous acid extracts of all L-streptococcal cultures studied reacted only with their specific antisera and did not cross-react with any other group specific streptococcal antigens. All 33 L-streptococcal cultures, in contrast to A-streptococci, produced beta-D-glucuronidase and beta-D-galactosidase, hydrolyzed Na-hippurate, grew on 10% and 40% bile blood agar and were agglutinated by the lectin of Arachis hypogaea. Some differences between A- and L-streptococci were also observed in their sensitivity patterns to bacitracin and sulfamethoxazole-trimethoprim.

Agglutination↗
PubMed1995

The proximal straight tubule (PST) basolateral cell membrane water channel: selectivity characteristics.

Proximal straight tubules (PST) were dissected from rabbit kidneys, held by crimping pipettes in a chamber and bathed in a buffered isosmotic (295 mOsm/kg) solution containing 200 mM mannitol (MBS). Changes in tubule diameter were monitored on line with an inverted microscope, TV camera and image processor. The PST were then challenged for 20 sec with MBS made 35 mOsm/kg hyperosmotic by addition of either NaCl, KCl, mannitol (M), glycerol (G), ethylene glycol (E), glycine (g), urea (U), acetamide (A) or formamide (F). With NaCl, KCl, M, G, E, g, U, and A, tubules shrunk osmometrically within 0.5 sec and remained shrunk for as long as 20 sec without recovering their original volume (sometimes A showed some recovery). PST barely shrunk with F and quickly recovered their original volume. The permeability coefficients were 0 microns/sec (NaCl, M, g, E and U), 1 micron/sec (A), 84 microns/sec (F) and 0.02 micron/sec (G). The reflection coefficients sigma = 1.0 (NaCl, KCl, M, G, E, g and U), 0.95 (A) and 0.62 (F). Similar sigma values were obtained by substituting 200 mOsm/kg M in MBS by either NaCl, KCl, G, E, g, U, a or F. The olive oil/water partition coefficients are 5 (M), 15 (U), 85 (A) and 75 (F) (all x 10(-5)). Thus, part of F permeates the cell membrane through the lipid bilayer. The probing molecules van der Waals diameters are 7.4 x 8.2 x 12.0 (M), 3.6 x 5.2 x 5.4 (U), 3.8 x 5.2 x 5.4 (A) and (3.4 x 4.5 x 5.4 (F) A.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1979

Effect of intralesional and systemic BCG-application or a combined cyclophosphamide/BCG treatment on experimental bladder cancer.

Infiltrating transitional cell carcinomas of the urinary bladder were induced by ingestion of 0.188% N-[4-(5-nitro-2-furyl)-2-thiazolyl] formamide (FANFT) in 145 female Wistar rats. After 8 months of carcinogen exposure, the animals were divided into different treatment groups. They received cyclophosphamide intraperitoneally as a single injection or BCG either once intralesionally or weekly subcutaneously or a combination of cyclophosphamide followed by subcutaneous BCG. The treatment effect was determined by body weight measurements and bladder tumour weight after 12 months. Compared with a control group statistically significant differences of bladder tumour weights were found after treatment with BCG alone or in combination with cyclophosphamide. Intralesional BCG resulted in an insignificant increase of tumour weights.

Animals↗
PubMed1982

Effect of partial cystectomy on the induction of pre-neoplastic lesions in rat bladder initiated with N-butyl-N-(4-hydroxybutyl)nitrosamine followed by bladder carcinogens and promoters.

The effect of partial cystectomy on the occurrence of pre-neoplastic lesions, papillary or nodular hyperplasia (PN hyperplasia), of the bladder in male F344 rats was studied in an experiment in which bladder carcinogens and promoters were given to the rats after initiation with BBN. The bladder carcinogens tested were N-ethyl-N-(4-hydroxybutyl)nitrosamine (EHBN) and N-4[4-(5-nitro-2-furyl)-2-thiazolyl]formamide (FANFT) and the bladder promoters were sodium saccharin, sodium cyclamate, and DL-tryptophan. Partial cystectomy significantly decreased the occurrence of PN hyperplasia in rats treated with EHBN and tended to inhibit that in rats given saccharin or tryptophan. Thus partial cystectomy inhibited rather than enhanced the induction of PN hyperplasia.

Animals↗
PubMed1979

The mechanism of C-binding: depurination and beta-elimination.

C-banding of chromosomes involves the differential solubilization of fragmented DNA from euchromatin by three sequential treatments: 1. Acid, 2. Mild base, 3. Hot salt. The data indicate solubilization is effected by 1) depurination, 2) DNA denaturation, 3) chain breakage of the depurinated sites respectively in the three treatments. Conditions were found wherein each treatment in proper sequence was necessary for C-banding and the appropriate chemical reactions were measured in these treatment conditions. The acid treatment (0.2 N HCl) depurinates chromosomal DNA at the rate of 0.26 x 10(-6) purines/dalton min to an alkaline molecular weight of 10(5) daltons but does not break the depurinated sites. Bleomycin can substitute for acid as a base removing agent. Sodium borohydride, by reducing the depurinated sugar's aldehyde thereby preventing chain breakage by the beta-elimination reaction, reversibly inhibits DNA-extraction. Chain breakage at the DNA's apurinic sites occurs not in the 2 min mild alkali treatment where the half-life for breakage is 26 min but in the 18 h hot salt treatment where the half-life for chain breakage is 1-2 h. Most of the DNA extraction occurs in the hot salt as 10(5) dalton fragments as measured in formamide gradients. Bleomycin is introduced as a substitute for HCl; it removes nitrogenous bases from DNA in situ while better preserving the morphology of the final C-banded chromosomes.

Animals↗
PubMed1976

The transcripts of Balbiani rings from Chironomus thummi. Giant RNA molecules with messenger characteristics.

Experimental results on the transcription of Balbiani rings BR 1 and BR 2 of Chironomus thummi salivary gland chromosomes are presented. The DNA of Balbiani ring 2, which is the most active puff in larvae, is transcribed into large RNA molecules of about 22 X 10(6) D which resist degradation by heating, formamide or urea treatment. The asymmetrical distribution of electrophoretic profiles of BR 2 RNA and the appearence of a symmetrical BR RNA peak in the nucleoplasm suggest the synthesis of (mainly) one RNA fraction in BR 2. The gel electrophoretic patterns of BR 1 RNA are, on the other hand, characterized by the appearance of two main fractions of high molecular weight RNA, one of which corresponds in molecular weight (about 22 X 10(6) D) to BR 2 RNA. The second RNA fraction is significantly smaller in molecular size (molecular weight: about 10 X 10(6) D) and, like the 22 X 10(6) D RNA fractions of the two Balbiani rings, resistant against heating in 8 M urea. Binding to poly (U) sepharose of a significant part of Balbiani ring RNA suggests the existence of poly (A) and/or oligo (A) sequences in the Balbiani ring RNA. -- In situ hybridization of BR RNA to the salivary gland chromosomes reveals accumulation of silver grains over the Balbiani ring regions only and demonstrates the restriction of BR DNA sequences to the corresponding Balbiani ring.

Animals↗
PubMed1991

Ultrastructural detection of DNA-incorporated bromodeoxyuridine in resin embedded brain tissue.

We describe a protocol for the ultrastructural detection of DNA-incorporated bromodeoxyuridine (BUdR) in resin embedded tissue by means of post-embedding immunogold labeling. The paraventricular zone of rat embryos brains was dissected, fixed either in paraformaldehyde or glutaraldehyde, and embedded in LR White. BUdR gold labeling was only found when thin sections were pretreated with 4 N HCl. Other DNA denaturing agents, such as Na ethoxide, formamide, formic acid, heat or HCl at lower concentrations were ineffective. Very little difference in the degree of labeling was found depending on the fixation. This method can be applied to investigate the fine structure of replicating cells in other in vivo conditions, such as human tumors.

Animals↗
PubMed1975

In situ hybridization of ribosomal DNA labelled with 125iodine to metaphase and lampbrush chromosomes from newts.

Methods are described for in situ hybridization of ribosomal DNA from Xenopus laevis, labelled in vitro with 125iodine, to mitotic and lampbrush chromosomes from Triturus cristatus carnifex. The hybridization reaction was carried out in a mixture containing 50% formamide, 4 X SSC, 0.1 M KI, at 37 degrees C, or in 2 X SSC, 0.1 M KI at 65 degrees C. Autoradiographs of mitotic metaphases from 2 males showed labelling over the middle of the short arm of one chromosome IX in each metaphase. In some cases, a region near the end of a longer chromosome was also labelled. In a lampbrush preparations, labelling was confined to a region identified as about 53 units, near the middle of the short arm of both halves of bivalent IX. The usefulness of the technique and the significance of the labelling of only 1 of the 2 chromosomes IX in mitotic preparations are discussed.

Animals↗
PubMed1975

The molecular weights of yeast ribosomal precursor RNAs.

The molecular weights of the predominant rRNA precursors as well as those of 26-S and 17-S mature rRNA from Saccharomyces carlsbergensis were determined by polyacrylamide gel electrophoresis in the presence of formamide. Mature 26-S + 5.8-S rRNA was found to have a molecular weight of 1.24 X 10(6) while their immediate precursor, 29-S RNA, had a molecular weight of 1.52 X 10(6). Values of 0.70 X 10(6) and 0.82 X 10(6) were obtained for the molecular weights of mature 17-S rRNA and its 18-S precursor. Finally the 37-S precursor, common to both 29-S and 18-S RNA, was found to have a molecular weight of 2.80 X 10(6). Each precursor rRNA, therefore, contains extra sequences not found at the next stage of maturation.

Electrophoresis, Polyacrylamide Gel↗
PubMed1988

Quantitative estimation of increased vascular permeability in acute inflammatory reaction in the chicken skin.

An increase in vascular permeability was estimated quantitatively in acute inflammatory reaction in the chicken using passive cutaneous anaphylaxis as an experimental model. Dye exuded in the cutaneous lesion was extracted by formamide and measured spectrophotometrically. The technique, though time-consuming, worked well in the chicken. The results suggested that the method can be profitably utilised in studies relating to avian inflammation.

Animals↗
PubMed1975

Circular ribosomal DNA and ribosomal DNA: replication in somatic amphibian cells.

Pulse labelled rDNA from cultured somatic cells of Xenopus laevis was examined by electron microscope autoradiography. The pattern of replication closely resembles that of bulk chromosomal DNA and differs considerably from rDNA synthesis during amplification in the oocyte. - About 0.15% of the rDNA molecules in the purified preparations were circular. The presence of interlocked circles of equal size indicates that the circles are not in vitro cyclization artefacts, but may represent free rRNA genes. A low frequency of circles was also seen in Xenopus blood rDNA. Their stability in high concentration of formamide suggests that they too did not arise after DNA extraction.

Animals↗
PubMed1978

Chemical composition of the peptidoglycan-free cell walls of methanogenic bacteria.

Cell walls were prepared from freeze-dried samples of 7 strains of Methanobacterium by mechanical disintegration of the cells followed by incubation with trypsin. Electron microscopy revealed the presence of sacculi exhibiting the shape of the original cells, on which no surface structure could be detected. Ultrathin sections of the isolated sacculi showed a homogenously electron dense layer of about 10--15 nm in width. The ash content varied between 8 and 18% of dry weight. The sacculi of all the strains contained Lys: Ala:Glu:GlcNAc or GalNAc in a molar ratio of about 1:1.2:2:1. In one strain (M. ruminantium M1) alanine is replaced by threonine, however, Neutral sugars and--in some strains--additional amounts of the amino sugars were present in variable amounts, and could be removed by formamide extraction or HF treatment without destroying the sacculi. No muramic acid or D-amino acids typical of peptidoglycan were found. Therefore, the sacculi of the methanobacteria consist of a different polymer containing a set of three L-amino acids and one N-acetylated amino sugar. From cells of Methanospirillum hungatii no sacculi, but tube-like sheaths could be isolated, which tend to fracture perpendicularly to the long axis of the sheath along the fibrills seen on the surface. The sheaths consist of protein containing 18 amino acids and small amounts of neutral sugars. They are resistent to the proteinases tested and are not disintegrated by boiling in 2% sodium dodecylsulfate for 30 min. The three Gram-negative strains Black Sea isolate JR-1, Cariaco isolate JR-1 and Methanobacterium mobile do not contain a rigid sacculus, but merely a SDS-sensitive surface layer composed of regularly arranged protein subunits. This evidence indicates that, within the methanogens, different cell wall polymers characteristic of particular groups of organisms may have evolved during evolution, and supports the hypothesis that the evolution of the methanogens was separated from that of the peptidoglycan-containing procaryotic organisms at a very early stage.

Acetates↗
PubMed1990

Microbial degradation of acrylamide monomer.

Acrylamide, a neurotoxic monomer with extensive industrial applications was found to be degraded by the microorganisms present in a tropical garden soil. A bacterium capable of degrading acrylamide was isolated from this soil by enrichment. It was found to be aerobic, gram-negative, motile, short rod and identified as Pseudomonas sp. The bacterium degraded high concentrations of acrylamide (4 g/l) to acrylic acid and ammonia which were utilized as sole carbon and nitrogen source for growth. An amidase was involved in the hydrolysis of acrylamide, which could act on other short chain amides like formamide and acetamide but not on acrylamide analogues: methacrylamide and N,N-methylene bisacrylamide. The enzyme was sensitive to catabolite repression by succinate both in presence as well as absence of nitrogen source.

Acrylamides↗
PubMed1979

Characterization of 10S RNA: a new stable rna molecule from Escherichia coli.

When cells of Escherichia coli are labeled with 32Pi for long periods of time and the cell content is subjected to electrophoresis in polyacrylamide gels, an RNA band appears which is about 10S in size. This band seems to contain three conformers. After treatment with formamide only a single band appears in this region of the gel, which contains 550 nucleotides as determined from its mobility. The complexity of the fingerprint of this material, after digestion with T1-RNase, is in agreement with the size as determined by the mobility, this confirming that indeed it is a single molecule. Composition of the T1-oligonucleotides was determined by digesting the T1-generated oligonucleotides with pancreatic RNase and T2-RNase. The quantitative and qualitative analysis of these digestions suggest that 10S RNA contains 609 nucleotides. The molecule contains, besides the four regular bases, one copy per molecule of the modified base pseudouridine. 10S RNA cannot be processed by cell extracts to tRNA-sized molecules and does not bind significantly to ribosomes, hence it is unlikely to be a tRNA precursor or an mRNA.

Base Sequence↗
PubMed1978

Simultaneous observation of quinacrine bands and silver grains on radiolabeled metaphase chromosomes.

A procedure is described for quinacrine banding of radiolabeled metaphase chromosomes for autoradiography. The chromosomes can be labeled either in vivo or by in situ hybridization. The banding procedure involves treating the slides with RNase and formamide and staining in quinacrine. The slides are then processed for autoradiography. After development of the photoemulsion, the chromosomes can be karyotyped with UV light by their fluorescent banding patterns and the silver grains overlaying the chromosomes can be demonstrated by the addition of tungsten light. It is possible by careful manipulation of the visible light to simultaneously observe both fluorescent bands and silver grains. This technique should significantly increase the accuracy of chromosome identification after autoradiography and decrease the time and effort required for such analysis.

Animals↗
PubMed1987

An electrophysiological study of skeletal muscle fibres in the 'muscular dysgenesis' mutation of the mouse.

Experiments were performed on muscles of 18-19 day mice fetuses affected with muscular dysgenesis (mdg). Action potentials generated by electrical stimulation or potassium depolarization failed to trigger muscle contraction in mdg muscle fibres. By contrast, muscle contraction could be obtained by caffeine (15 mM) and, to a lesser degree, by nerve stimulation. We conclude that a defect in excitation-contraction (E-C) coupling is the cause of muscle paralysis. An early after potential (EAP) was present in the decay phase of the action potential and a potential 'creep' occurred in response to hyperpolarizing current pulses which can be taken as evidence for the presence of T-tubules in mdg muscle fibres. Data obtained from square pulse analysis and EAP measurements indicate larger input impedance and membrane time constant in mdg as compared to controls, which contrasts with similar surface membrane time constant (as estimated from the foot of the action potential) in both types of muscle. The excitability of the T-tubule system was tested by recording action potentials at early stages of TTX (5 X 10(-7) M) perfusion or washout in mdg and control muscles. In both cases, the action potentials decreased in amplitude and rate of rise and displayed two peaks, the second of which was suppressed by detubulation using the formamide treatment. This indicates action potential generation in the T-tubule membrane of mdg muscles. In all the impaled muscle fibers, nerve stimulation evoked epps which were accompanied by a weak local contraction in relation with Ca2+ influx through postsynaptic channels.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗
PubMed1979

A quantitative cytochemical method for the demonstration of delta5,3beta-hydroxysteroid dehydrogenase activity in unfixed tissue sections of rat ovary.

A method for the quantitative measurement of delta5,3beta-hydroxysteroid dehydrogenase activity in unfixed tissue sections of rat ovary has been described. The method depends on the oxidation of dehydroepiandrosterone (DHEA) and uses nitroblue tetrazolium as the final electron acceptor. Although the dehydrogenase is not a soluble enzyme, polyvinyl alcohol is included in the reaction medium to allow the use of a high substrate concentration whilst employing a low concentration (5%) of dimethyl formamide. The enzyme is equally dependent on NAD+ or NADP+ for its activity and this activity is significantly enhanced by the presence of cyanide. The NADP+ dependence is not abolished by inhibiting nonspecific alkaline phomonoesterase. The activity of delta5,3beta-hydroxysteroid dehydrogenase is completely dependent on a functional sulphydryl group. Furthermore, the enzyme activity is totally inhibited in the presence of a steroid substrate analogue at 10(-4) M.

3-Hydroxysteroid Dehydrogenases↗
PubMed1985

Immunocytochemical studies using a monoclonal antibody to bovine cardiac titin on intact and extracted myofibrils.

A monoclonal antibody specific to bovine cardiac titin has been identified. The antibody recognizes a common antigenic site in striated muscles of several species. In relaxed myofibrils, specific staining at the A-I junction resulted in a doublet of fluorescent bands within a sarcomere. The distance between the doublets in successive sarcomeres varied according to the degree of myofibrillar contraction. Staining on formamide-extracted myofibrils has confirmed that this epitope is located near the outer edges of isolated A bands. Selective extraction of myofibrillar proteins resulted in different staining patterns. Disrupting the structural integrity of the M-line or the A-band centre caused a significant amount of titin to translocate toward the Z-line region. In contrast, shortening of the A-band by removal of myosin from the ends of the thick filaments resulted in anti-titin staining moving closer to the M-line region. Several conclusions can be drawn from this study: (a) two aligned groups of titin molecules are placed symmetrically to the M-line in a sarcomere; (b) titin may attach directly or via intermediary protein(s) to sites near the M-line and Z-line such that the protein is under tension and (c) removal of proteins from either region results in titin staining in the opposite region. However, the edges of the A-band give some hindrance to collapse of the titin toward the M-line.

Animals↗