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Biomedical subjects

M Reuter

Publications and source records attributed to M Reuter.

At least 199 records · Page 11Linked to original sources

DNA methylation of bacterial viruses T3 and T7 by different DNA methylases in Escherichia coli K12 cells.

We have investigated the susceptibility of the genomes of the related bacteriophages T3 and T7 to the three major DNA methyltransferases (EcoK, dam, dcm) of their host, Escherichia coli K12. In vivo the EcoK host specificity enzyme only methylates the DNA of ocr- phages. This is due to an inhibition of the enzyme by the phage ocr+ gene product, which had previously been shown to be an inhibitor of the restriction endonuclease. EcoK-specific DNA methylation protects the ocr- viruses after one growth cycle on these host cells against the action of corresponding restriction endonuclease EcoK. Owing to the unique S-adenosyl-L-methionine hydrolase (sam+) activity of the T3-coded ocr+ protein, the T3 DNA is absolutely devoid of the methylated bases 6-methylaminopurine and 5-methylcytosine. In contrast to this, T7 derivatives and sam- derivatives of T3 carry a small number of about 2-4 molecules 6-methylaminopurine and 5-methylcytosine per genome. The presence of 6-methylaminopurine is due to dam methylation, though the majority of dam sites remain unmethylated. In vivo as well as in vitro the ocr+ protein has no influence on the activities of the dam and dcm methylase. The experiments gave some evidence for the existence of a second cytosine methylase in E. coli K12. Besides dam and dcm recognition sites being undermethylated, their absolute number in T3 and T7 DNAs is far below the expected value. Moreover, one of the two dcm sites present in T7 (Studier strain) is missing in our T7 strain owing to a 1300-base-pair deletion in gene 0.7.

5-Methylcytosine↗

Neuropeptides in a microturbellarian--whole mount immunocytochemistry.

A whole mount immunofluorescence method was used for the localization of immunoreactivity (IR) to four regulatory peptides and the bioamine serotonin in the nervous system of Stenostomum leucops (Turbellaria, Platyhelminthes). The flatworm S. leucops belongs to the taxon Catenulida which, according to the new phylogenetic system by Ax [2], forms a key group between the coelenterates and more advanced flatworm species. Positive IR was obtained using antisera against FMRF-amide, beta-endorphin, growth hormone releasing factor (GRF), substance P, and serotonin. The distribution patterns of these neuropeptide-like immunoreactivities differ significantly from each other. Antisera against Leu-enkephalin, bovine pancreatic polypeptide (BPP), bombesin, cholecystokinin (CCK-8), neurotensin, somatostatin, growth hormone (GH), secretin, and neurophysin II gave negative results. This primitive flatworm shows similarities with hydra in the lack of IR to anti-somatostatin, anti-Leu-enkephalin, and anti-BPP. These antisera give positive IR in more advanced flatworm species, indicating a later convergent evolution of vertebrate-like peptides within the phylum Platyhelminthes.

Animals↗

Immunocytochemical demonstration of peptidergic neurons in the central and peripheral nervous systems of the flatworm Microstomum lineare with antiserum to FMRF-amide.

The central nervous system (CNS) and the peripheral nervous system (PNS) of the flatworm Microstomum lineare were studied by means of the peroxidase-antiperoxidase (PAP) immunocytochemical method, with the use of antisera to the molluscan cardioactive peptide FMRF-amide. FMRF-amide immunoreactive perikarya and nerve fibres are observed in the CNS and the PNS. In the CNS, immunoreactive perikarya and nerve fibres occur in the brain, in the epithelial lining and the mesenchymal surroundings of the ciliated pits, and positive fibres in the longitudinal nerve cords. In the PNS, immunoreactive fibre bundles with variocosities occur in the pharyngeal nerve ring, in symmetrical groups of perikarya on each side of the pharynx, and in the mouth area. Positive perikarya and meandering nerve fibres appear in the intestinal wall. A few immunoreactive cells and short nerve processes are observed at the male copulatory organ and on both sides of the vagina. Some immunoreactive peptidergic cells do not correspond to cells previously identified by histological techniques for neurosecretory cells. The distribution of immunoreactivity suggests that the FMRF-amide-like substance in CNS and PNS in this worm has roles similar to those of the brain-gut peptides in vertebrates. The status of FMRF-amide-like peptides as representatives of an evolutionarily old family of peptides is confirmed by the positive immunoreaction to anti-FMRF-amide in this primitive microturbellarian.

Animals↗

DNA methylation of T3 virus ocr+ and ocr- strains in Escherichia coli cells harbouring the EcoK DNA host specificity system.

The influence of the T3 gene functions ocr+ and sam+ on the extent of phage DNA methylation in Escherichia coli K12 cells was studied by determining the proportion of 6- methylaminopurin to adenine in the purified DNA of T3 wild-type, sam- and ocr- sam- phage strains. We demonstrate that the DNA of T3 ocr- sam- mutants carries 12 methyl groups as a result of the action of the host-specificity methylase EcoK . In contrast to this the DNA of ocr+ strains is not EcoK -specifically methylated.

DNA, Bacterial↗

Restriction of bacteriophage T3 and T7 ocr+ strains by the type II restriction endonuclease EcoRV.

When E. coli cells carrying the plasmid pLG13 (coding for the newly discovered type II restriction endonuclease EcoRV) are infected with phage T3 or T7, only T7 is able to replicate normally. T3 wild-type as well as its ocr- mutants are subject to DNA restriction in vivo and in vitro. The EcoRV enzyme cuts T3 DNA at 5 sites. T7 and its ocr- mutants have no EcoRV sites in their DNA. In contrast to the anti-restriction activity of the T3 and T7 ocr+ gene function against type I and III restriction enzymes, the ocr+ protein is unable to inactivate the type II restriction endonuclease EcoRV.

DNA Replication↗

Influence of phage T3 and T7 gene functions on a type III(EcoP1) DNA restriction-modification system in vivo.

The ocr+ gene function (gp 0.3) of bacteriophages T3 and T7 not only counteracts type I (EcoB, EcoK) but also type III restriction endonucleases (EcoP1). Despite the presence of recognition sites, phage DNA as well as simultaneously introduced plasmid DNA are protected by ocr+ expression against both the endonucleolytic and the methylating activities of the EcoP1 enzyme. Nevertheless, the EcoP1 protein causes the exclusion of T3 and T7 in P1-lysogenic cells, apparently by exerting a repressor-like effect on phage gene expression. T3 which induces an S-adenosylmethionine hydrolase is less susceptible to the repressor effect of the SAM-stimulated EcoP1 enzyme. The abundance of EcoP1 recognition sites in the T7 genome is explained by their near identity with the T7 DNA primase recognition site.

DNA Replication↗

Changes in angiotensin-converting enzyme activity and angiotensin I level in asthmatic and healthy children after submaximal physical work.

The changes in angiotensin-converting enzyme activity and serum angiotensin I levels have been studied in 16 controls subjected to submaximal physical work. Baseline (Pre-exercise) angiotensin I levels were identical in both groups. Physical exercise caused an elevation that was more marked in the asthmatic group than in the control group. The activity of serum angiotensin-converting enzyme differed in the two groups even before physical exercise, the asthmatic children having exhibited an activity level significantly lower than that of the healthy controls. after submaximal work, the enzyme activity increased in healthy subjects but decreased in asthmatic children.

Angiotensin I↗

The nervous system of Microstomum lineare (Turbellaria, Macrostomida). II. The ultrastructure of synapses and neurosecretory release sites.

The ultrastructure of release sites of neurochemical messenger substances in the microturbellarian Microstomum lineare was examined. Aminergic neurites form conventional synapses and synapse-like structures (SLS). Variants of true synapses include: "single" synapses with symmetric pre- and postsynaptic densities, "shared" synapses, i.e., contacts between 1 pre- and 2 postsynaptic fibres, en passant synapses between parallel axonal membranes, and synapses without thickenings having only clustered vesicles in the presynaptic terminal. SLS on a nerve cell soma or facing an intercellular stromal channel near muscles are described. Peptidergic neurites containing large granular vesicles (LGV) from synaptoids and signs of putative neurosecretory release. Synaptoids between neurites and between neurite and muscle have lucent vacuoles (about 100 nm) and dense material at the contact site. In en passage synaptoids dense-core vesicles are embedded in electron-dense material at the contact site. Putative signs of release of neurosecretory material other than "typical" exocytosis have been observed.

Amines↗

[Protection of foreign DNA against host-controlled restriction in bacterial cells. II. Protection of pSF2124 plasmid by the gene function of bacteriophages T3 and T7].

When restriction-active Escherichia coli cells (R+P1m+P1) are transformed with the pSF2124 plasmid, a common vector in experimental gene transfer, the efficiency of transformation (e.o.t.) is lowered by 2 orders of magnitude compared with restriction-negative (r-P1m-P1 or r-P1m+P1) recipient cells due to restriction of the pSF2124 DNA by endoR.EcoP1. Preinfection of r+P1m+P1 cells with UV-inactivated ocr+ phages (T3, T7, T3sam-) still able to express their early genes protects the plasmid DNA against restriction by endoR.EcoP1: The e.o.t. of r+P1m+P1 recipient cells with pSF2124 attains the same high value as that of r-m- cells. The specific role of the ocr+ gene function was demonstrated by the use of ocr- mutants (T3/R7, T7/D111): Preinfection with such phage mutants does not increase the e.o.t. of r+P1m+P1 cells. An unspecific e.o.t. alteration of restriction-negative (r-m-) recipient cells by ocr+ or ocr- phages was excluded. The ocr+ gene function can be exploited to protect pSF2124 against DNA restriction. The recipient cells survive the process of phage preinfection and transformation and stably replicate themselves as well as the plasmid DNA.

DNA Restriction Enzymes↗

Ultrastructure of epidermal eyespots of Microstomum lineare (Turbellaria, Macrostomida).

The eyespots of Microstomum lineare were studied by electron microscopy, light microscopy, and fluorescence microscopy. Each eyespot consists of two ciliary photoreceptor cells shielded by pigment cells and additional sensory cells. The photoreceptor cells are characterized by a distal intracellular cavity lined with 50-100 interwoven cilia. The other sensory cells are of two ultrastructurally different types, one with long cilia predominating and the other with balloonlike cilia. The pigment cells, which envelop processes of the sensory cells, contain pigment vacuoles varying in size and content and give a bright red fluorescence by the Falck-Hillarp method. The eyespots are suggested to perform a dual function as photoreceptors and chemoreceptors. The evolutionary significance of ciliary photoreceptors in Turbellaria is discussed.

Animals↗

The nervous system of Microstromum lineare (Turbellaria, Macrostomida). I. A fluorescence and electron microscopic study.

The nervous system (NS) of Microstomum lineare (Turbellaria, Macrostomida) was studied by electron and light microscopy, combined with fluorescence histochemistry (Falck-Hillarp method for biogenic monoamines). The NS is primitively organized, with a bilobed brain, two lateral nerve cords lacking commissures, and peripheral nerve cells scattered along the nerve cords. The stomatogastric NS, with a pharyngeal nerve ring, is joined to the central NS by a pair of connective ganglia. A green fluorescence in all parts of the NS indicates catecholaminergic neurons as the dominant neuron type. Ultrastructurally, two types of neurons were identified on the basis of their vesicle content: 1. Aminergic (catecholaminergic) neurons containing dense-core vesicles of varying electron-density and size, i.e., small dense-core vesicles (diameter 50--100 nm), vesicles with a highly electron-dense core (60--140 nm), and vesicles with an eccentric dense-core. 2. Presumed peptidergic neurosecretory neurons containing large granular vesicles (diameter about 200 nm) in the stomatogastric NS and peripheral parts of the central NS. In light microscopy, paraldehyde-thionin stained neurons were observed in the same areas.

Animals↗

[20 years of prostate surgery. Reduction in the number of complications through application of a new concept in TUR, cryosurgery and surgical procedures].

During the last 20 years a deep change took place in the technique as well as in the conception of the operations of the prostate gland in adenomas and carcinomas. The endoscopic methods were adapted to the physiology of the urinary tract and to the condition of the patient; thus the general complication and lethality rate was decisively lowered (reduction of the general postoperative lethality from 1.2% to 0.77%). -- The operation method is selected according to the degree of risk of the patient. The TURP with the new low pressure technique (72%) and the freezing (25%) also in the adenoma of supersize are preferred. On the other hand, the surgical interventions rich in loss (lethality 3.3-4.3%) are decisively reduced (below 3%). -- The new endoscopic technique of cryosurgery guarantees a high value operation also for patients with a high risk (lethality 1.8%), in which cases the failure rate of the other method is indirectly decisively reduced (TURP 0.3%, radical TUR 0.7%). The carcinoma is (sub-) radically resected, and resected with increased risk or relapse, respectively; lymphadenectomy and radiation therapy supplement these operations.

Adenoma↗