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M K Wolf

Publications and source records attributed to M K Wolf.

At least 55 records · Page 3Linked to original sources
PubMed1988

Shiverer jimpy double mutant mice. IV. Five combinations of allelic mutations produce three morphological phenotypes.

Mice which simultaneously express mutant genes at both the shi and jp loci (double mutant mice) have phenotypes much more complex than simple addition of individual mutant characteristics. Morphological study of shi jpmsd, shimld jp, and shi/shimld jp, and comparison with previously studied shi jp and shimld jpmsd, shows that 3 classes of central nervous system (CNS) white matter morphology are produced. (1) shi jp shows suppression of most jp characteristics: it is like shi except for more major dense line, and possibly more myelin, than shi alone. No other combination has as much myelin or any major dense line at all. (2) shi jpmsd has qualitative and quantitative characteristics intermediate between the two single mutants. (3) All combinations studied involving shimld have much less myelin than either single mutant. Qualitatively they express most jp locus features but suppress all shi locus features except abnormalities of myelin compaction. The difference between shi and shimld has more influence on the double mutant morphology than the difference between jp and jpmsd. In the 3-mutant combination, shi/shimld jp, the influence of shimld completely overrides that of shi. These morphological phenotypes resist assignment to any hierarchy of normalcy, and their specific features have no simple explanation in presently known molecular biology of the shi and jp locus mutations. They suggest the possibility of multiple copies and multiple primary functions of the messages at these loci.

Animals↗
PubMed1988

Correlation between verapamil plasma concentration and P-R prolongation in essential hypertension.

Plasma verapamil concentration was correlated with serial electrocardiographic P-R intervals in patients with essential hypertension receiving immediate-release (80 to 120 mg three times a day) or sustained-release (240 mg daily) verapamil. The mean P-R interval in 22 patients taking placebo and immediate-release verapamil was 0.18 second. The borderline first-degree atrioventricular block of three patients did not change during treatment. Plasma verapamil concentrations of patients with a P-R interval longer than 0.20 second and of those with a P-R interval of 0.20 second or less were 169 +/- 73 ng/mL and 63 +/- 8 ng/mL, respectively. Six patients taking sustained-release verapamil had a maximal mean P-R interval of 0.19 +/- 0.01 second during 24-hour ambulatory electrocardiographic monitoring. P-R intervals were 0.22 second or more in two patients, but they returned to normal by hour 7 for one and by hour 20 for the other patient. In summary, transient P-R prolongation occurred with oral verapamil therapy, but no patient, regardless of baseline P-R interval, developed high-grade atrioventricular block.

Delayed-Action Preparations↗
PubMed1988

Characterization of the plasmid from Escherichia coli RDEC-1 that mediates expression of adhesin AF/R1 and evidence that AF/R1 pili promote but are not essential for enteropathogenic disease.

RDEC-1, an Escherichia coli strain that adheres to rabbit mucosa and causes an attaching, effacing lesion, expresses the pilus adhesin AF/R1 which determines in vitro attachment to rabbit intestinal brush borders. In order to determine the role of AF/R1 pili in the pathogenesis of enteropathogenic diarrhea in rabbits, we localized the genes for AF/R1 expression, constructed an AF/R1- strain, and compared the virulence of the AF/R1+ and AF/R1- strains with particular attention to the development of attaching, effacing lesions. We introduced Tn5 into the 86-megadalton (MDa) conjugative plasmid known to mediate expression of AF/R1 pili and transferred the derivative plasmids into laboratory strain HB101. Transconjugant M5 was found to contain the 86-MDa plasmid from RDEC-1 and to express AF/R1 pili. Pilus expression on M5 was confirmed by reaction with antiserum raised against purified AF/R1 pili and allowed the bacteria to adhere to the rabbit ileum in an in vitro assay. Three Tn5 insertions in the 86-MDa plasmid were obtained which resulted in loss of AF/R1 expression. Part of the plasmid was mapped, including a region necessary for AF/R1 pilus expression. AF/R1- mutant strain M34 was constructed, and its pathogenesis was investigated. M34 produced disease in rabbits but was less virulent than the parent. The characteristic effacing lesions of RDEC-1 and enteropathogenic E. coli developed in the intestine of rabbits infected with either M34 or RDEC-1, although with M34 they were much less frequent and did not involve the small bowel. We conclude that AF/R1 pilus expression is not essential for the attaching, effacing lesion but serves as an accessory virulence factor which promotes an initial interaction of RDEC-1 with normal epithelial cells.

Adhesins, Escherichia coli↗
PubMed1988

New aerobactin-mediated iron uptake system in a septicemia-causing strain of Enterobacter cloacae.

Unlike the great majority of the aerobactin-producing enteric bacteria documented in the literature, Enterobacter cloacae EK33, isolated from a case of human neonatal meningitis, did not show any homology at the DNA level with the prototype aerobactin system encoded by the ColV-K30 plasmid. However, both the nuclear magnetic resonance spectrum and fast-atom bombardment mass spectrometry of the siderophore purified from EK33 confirmed its identity with aerobactin. Bioassay screening of a gene library of total DNA of EK33 led to the isolation of several aerobactin-positive clones. Under conditions of iron limitation, these clones expressed in Escherichia coli a protein of 72 kilodaltons that reacted with antiserum raised against the pColV-K30 74-kilodalton aerobactin receptor, while the original E. cloacae strain synthesized an 85-kilodalton protein which also cross-reacted with the antiserum. Restriction endonuclease analysis of the cloned DNA confirmed the structural differences between the two aerobactin genetic systems.

Conjugation, Genetic↗
PubMed1987

Shiverer jimpy double mutant mice. III. Comparison of shimld*jpmsd and shi*jp phenotypes demonstrates dissimilar interactions of allelic mutations.

Double mutant mice, which are of the genotype shimld/shimld*jpmsd/Y and therefore express both the shimld and jpmsd mutations, have a CNS myelin protein composition which resembles shimld/shimld alone but not jpmsd/Y alone. The double mutant CNS white matter morphology shows much less myelin and major dense line than either shimld/shimld or jpmsd/Y, but has other features which resemble jpmsd/Y but not shimld/shimld. In contrast, the parallel double mutant shi/shi*jp/Y, which expresses the alleles shi and jp rather than shimld and jpmsd, has already been shown to have biochemical and morphological phenotypes which are consistent with each other, both being intermediate between shi/shi and jp/Y and therefore suggesting partial reciprocal intergenic suppression (Brain Research, 374 (1986) 45-53 and 54-62). To assist in explaining the apparent inconsistencies between the biochemical and morphological phenotypes of the shimld/shimld*jpmsd/Y double mutant and between interactions of allelic mutations at the shi and jp loci, a hypothesis of multiple primary gene functions at these two loci is proposed.

Animals↗
PubMed1986

Shiverer jimpy double mutant mice. II. Morphological evidence supports reciprocal intergenic suppression.

Mice which carry both the shiverer (shi) and the jimpy (jp) mutations have a morphological phenotype with features of each single mutation by itself but in milder form: the number of myelin sheaths is increased relative to jp, the thickness of sheaths and amount of major dense line is increased relative to shi, and the abnormal, lipid-filled cells characteristic of jp are not seen. However, the abnormal bundles of oligodendrocyte microprocesses and errors in the targeting of myelination which characterize shi are not altered by the presence of the jp mutation. This morphological evidence suggests partial reciprocal intergenic suppression in shiverer jimpy double mutant mice and therefore agrees with conclusions based on biochemical data presented by Kerner and Carson (Brain Research, 374 (1986) 45-53).

Animals↗
PubMed1986

Genotype-specific myelin formation around normal axons in cytosine arabinoside-treated organotypic cultures injected with normal or shiverer optic nerve.

Organotypic cultures of newborn mouse cerebellum, treated with 'antimitotic' drugs to prevent myelination, contain abundant large neurons and axons but no mature oligodendroglia and no myelin. When oligodendroglia, contained in optic nerve fragments from 7-12-day-old normal or shiverer mutant mouse, are added to the cultures, myelin is formed in the vicinity of the optic nerve explant. The phenotype of the added myelin corresponds to the genotype of the optic nerve added, indicating that the added oligodendroglia are making the myelin themselves rather than contributing a diffusible factor which stimulates native cerebellar oligodendroglia or their precursors. This system will be valuable for direct, detailed comparisons of myelin formation by normal and mutant oligodendrocytes.

Animals↗
PubMed1986

Evidence for the role of a siderophore in promoting Vibrio anguillarum infections.

Vibrio anguillarum strain 775 harbouring the virulence plasmid pJM1 produces a plasmid-mediated siderophore that can crossfeed siderophore-deficient, receptor-proficient mutants of V. anguillarum in vitro. Experimental infections of salmonid fishes with mixtures consisting of the wild-type strain and a siderophore-deficient, receptor-proficient mutant resulted in recovery of both the wild-type and the mutant strain, while in infections with mixtures consisting of the wild-type strain and a siderophore-deficient, receptor-deficient mutant only the wild-type strain could be recovered. These results suggest that the V. anguillarum plasmid-mediated siderophore is produced in vivo in a diffusible form and that it is an important factor of virulence.

Animals↗
PubMed1986

Migration and myelination by adult glial cells: reconstructive analysis of tissue culture experiments.

Adult glia are capable of at least limited myelination of CNS axons. However, it is difficult to quantitate their myelination or migratory capacities and to examine contributions of the CNS environment or exogenous factors that might promote or inhibit this process in situ. We have therefore developed a mouse tissue culture system in which optic nerve glia (in the form of appropriately handled optic nerve) are added to chemically demyelinated cerebellar axons. Optic nerve up to postnatal day 411 (P411) contains cells that can migrate out of the nerve into the cerebellar explant and form myelin around its axons. The success rate for myelin formation in these cultures is 57% for immature (P7-11) glia and 55% for adult (P50-411) glia. Computer-generated reconstructions of cultures containing immature (P8) and adult (P89 and P139) nerves demonstrate that in all 3 cases the glia may migrate more than 0.6 mm before myelinating axons, assuming the shortest possible track. Both the age limit for myelination and distance limit for migration, if any, remain to be determined for these adult glia. In successful cultures, myelin always directly abuts the optic nerve surface, whether or not it also extends further, suggesting that migrating glia may depend upon contact guidance by myelin-receptive axons. We conclude that this culture system is a useful model of adult CNS myelin regeneration, in which one can examine the influence of potential trophic or toxic factors on specific aspects of myelinating glial cell behavior.

Animals↗
PubMed1985

Myelin deficient (shimld) mutant allele: morphological comparison with shiverer (shi) allele on a B6C3 mouse stock.

A new B6C3 stock of shimld mutant mice is compared in terms of behavior and CNS morphology with both a B6C3 shi stock and reports on other shimld animals. Defects of B6C3 shimld myelination seen at postnatal day 21 (P-21) are comparable to those in B6C3 shi with respect to % axons myelinated, sheath thickness, errors in the wrapping and targeting of myelin and abnormal oligodendrocyte shape. The two mutations are similarly expressed in cerebellar organotypic cultures. However, the major dense line (MDL) is present in a few shimld myelin sheaths at P-21 and a few sheaths show myelin basic protein by immunocytochemistry, while neither phenomenon is seen in shi at this age in the same CNS regions. Shimld mice survive their disease significantly better than shi. The shimld stock currently under study elsewhere differs from this B6C3 stock in that MDL was reported only in older animals, and behavior and survival were severely compromised.

Alleles↗
PubMed1984

Cultures of shiverer mutant cerebellum injected with normal oligodendrocytes make both normal and shiverer myelin.

Earlier reports suggested that injecting normal optic nerve into organotypic cerebellar cultures from three of the central nervous system hypomyelinated mutant mice resulted in striking local increases in myelination of mutant axons. It has been questioned whether this myelin was produced by the introduced normal glia because there was no "marker" by which the genotype of an individual myelin sheath could be rigorously determined. The present study shows that the myelin sheaths of the central nervous system hypomyelinated mutant shiverer (shi/shi) can be distinguished in vitro from genetically normal myelin by their immunocytochemical reactions and ultrastructure. When shi/shi cultures are injected with normal optic nerve, they produce two kinds of myelin as distinguished by these techniques: shi/shi myelin throughout and, in addition, ultrastructurally and immunocytochemically normal myelin near the optic nerve. These results suggest that the primary defect of the shi/shi mutation involves the oligodendrocyte and support the earlier conclusion that in all of the glial injection experiments to date the genotype of the oligodendrocyte determines the phenotype of the myelin produced.

Animals↗
PubMed1983

Hypomyelinated mutant mice. V. Relationship between jp and jpmsd re-examined on identical genetic backgrounds.

jp and jpmsd, two allelic mutations in the mouse that sharply reduce the amount of CNS myelin, produce diseases that can be distinguished morphologically only by their severity. This has raised the question of whether the two mutations are truly distinguishable. Since the two mutations have never been maintained on the same genetic background, correct quantitative and morphological comparison have not been possible. We have prepared a B6C3H stock of jp on the same genetic background as the available stock of jpmsd. In this jp stock, behavioral abnormalities, relative proportion of myelinated axons, and major morphological characteristics of the disease in situ are unchanged from the previous jp stock. The jp disease continues to be more severe than that of jpmsd. However, tissue from the new B6C3H stock myelinates better in organotypic culture than previous jp stocks. The increase in myelination is advantageous, not only for accurate comparison of the two alleles but for all culture studies of jp. Strictly comparable strains or stocks should be utilized in any comparative studies of closely related mutations such as jp and jpmsd.

Alleles↗
PubMed1983

Myelination of jp,jpmsd, and qk axons by normal glia in vitro: ultrastructural and autoradiographic evidence.

Normal optic nerve glia were 'injected' into hypomyelinated mutant jp,jpmsd, and qk cerebellum by co-culturing explants in direct physical contact. Quantitative light microscopic studies demonstrated that such glial injection significantly increased the number of myelin profiles counted in cultures, suggesting that axons in all 3 mutants can accept myelination from competent glia when they are made available. In each mutant, the observed increase in myelination was independent of the ages of donor optic nerves and recipient cultures, but absolutely required positioning of the optic nerve so that direct contact occurred with the mutant cerebellar explants. The additional myelin found near the zone of fusion with the optic nerve morphologically resembled normal, not mutant myelin. Autoradiographs made after [3H]thymidine-labeled normal optic nerve was injected into jpmsd cultures showed that labeled cells had colonized the nearby mutant tissue. Labeled cells identified as oligodendrocytes by ultrastructural criteria were found adjacent to myelin segments near the fusion zone, but direct continuity between processes of these oligodendrocytes and myelin sheaths was not demonstrated. The astrocytes and phagocytic cells which were also labeled had no obvious relationship to myelinated axons. These results provide experimental evidence that the primary abnormalities produced by the three mutations jp,jpmsd, and qk are inherent in their glial cells, probably although not definitely in the oligodendrocytes.

Animals↗
PubMed1981

Increased binding of a hydrophobic, photolabile probe to Escherichia coli inversely correlates to membrane potential but not adenosine 5'-triphosphate levels.

We describe conditions for a quantitative determination of azidopyrene binding to Escherichia coli cells. In addition, we define conditions whereby irradiation of azidopyrene in the presence of cells leads to irreversible association of probe with cells. This is presumably due to the light-dependent generation of reactive nitrenes and subsequent incorporation of nitrenopyrene moieties into cellular components. These methods allowed us to determine that the amount of azidopyrene bound to cells was inversely correlated with the magnitude of the cellular membrane potential, but was not correlated with high or low adenosine 5-triphosphate levels per se. Cells bound more azidopyrene if the delta psi was low. Cell-bound azidopyrene was found to be entirely associated with the inner and outer membrane. We suggest that the decreased association of hydrophobic probes upon energization of whole cells reflects a rapid transition in structural properties of the cell envelope.

Adenosine Triphosphate↗
PubMed1980

Hypomyelinated mutant mice. II. Myelination in vitro.

Organotypic cultures of cerebellum from hypomyelinated mutant mice provide a powerful experimental system for studying the cell biology of the mutant diseases. We have examined the extent to which the culture system reproduces the diseases of three well-known mutants, qk, jpmsd, and jp. Quantitation of myelin profiles per sq. mm of section demonstrates that in vitro, as in situ, qk produces the most myelin jpmsd an intermediate amount, and jp the least. Myelin in qk cultures is unique in being invisible by light microscopy of the living culture. Hypomyelination of jp may be more severe in vitro than in situ. Cultures of jpmsd exhibit many of the ultrastructural features of cerebellar abnormalities that occur in situ: degree of hypomyelination, clustering of myelin segments, scarcity of oligodendrocytes, absence of nodes of Ranvier but presence of heminodes, and apparent structural integrity of the myelin sheaths. Correspondence between in vitro and in situ ultrastructure is more difficult to assess for jp, because the available sample of jp myelin in vitro is too small, and for qk, because the abnormalities observed in situ resemble nonspecific abnormalities of normal myelin in vitro.

Animals↗
PubMed1980

Hypomyelinated mutant mice: description of jpmsd and comparison with jp and qk on their present genetic backgrounds.

Hypomyelinated mutant mice are valuable natural animal systems for analysis of CNS myelin development, chemistry and diseases. One mutant, jpmsd, has never received thorough morphological description. We here describe the detailed morphology of jpmsd, compare it with well-studied jp and qk on their present genetic backgrounds, and discuss the genetic histories of all 3 mutants. Region for region, jpmsd has twice as much myelin as jp, but 1/2--1/5 as much as qk. Both jp and jpmsd have scarce oligodendrocytes, rare nodes of Ranvier, clustering of myelin segments, abnormal lipid-filled cells, frequent degenerating cells, and rare distorted myelin profiles. In contrast, qk has abnormally numerous oligodendrocytes, frequent nodes of Ranvier, no obvious myelin clustering, no lipid-filled cells, rare degenerating cells, and frequent abnormal or distorted myelin profiles. jp and jpmsd are quantitatively different, but qualitatively similar. Since its origin, the jpmsd disease has inadvertently been ameliorated by transferring the mutation to a different background. Persistent differences in the remainder of the genome might account for all remaining apparent differences between jp and jpmsd. In contrast, qk is totally dissimilar in morphology and presumably in pathophysiology.

Animals↗