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Biomedical subjects

L K Miller

Publications and source records attributed to L K Miller.

At least 199 records · Page 11Linked to original sources

Newborn northern fur seals (Callorhinus ursinus)--do they suffer from cold?

Several hundred thousand northern fur seals (C. ursinus) are born each summer during July at St. Paul Island in the Bering Sea. The weather in the area is usually cold, wet, and windy during the breeding season. At birth the pups are small (5--6 kg) and insulated only by a partly wettable pelt and a 2- to 4-mm layer of blubber. In air, the pups' lower critical temperature appears to be below the 6 degrees C 50-yr record low July temperature for the islands. During rainy weather much of the insulative value of the pelt is lost, and the pups, which already have a high resting metabolic rate of 3.5 W.kg-1, must increase heat production by shivering and/or nonshivering thermogenesis to maintain deep body temperature. The high level of metabolism (up to 18 W.kg-1) is supported by a very rich milk. The pups will, nevertheless, become hypothermic if their insulation is not improved through peripheral vasoconstriction and shedding of water from the pelt by periodic shudder. Even with these protections the newborn and very young pups are brought close to their limit of tolerance during rainy and windy days. Unfit pups are likely to succumb under such circumstances.

Adipose Tissue↗

Restriction endonuclease analysis for the identification of baculovirus pesticides.

Gel electrophoresis of deoxyribonucleic acid (DNA) fragments generated by digesting the DNA genomes of nuclear polyhedrosis viruses (NPV) with restriction endonucleases provides DNA fragment patterns that may be used to identify different viruses of this group. Characteristic fragment patterns were obtained for three NPVs, which are important as biological pesticides (Autographa californica NPV, Orgyia pseudotsugata NPV, and Heliothis zea NPV). The DNA fragment patterns of the A. californica NPV genoms did not change with passage through the alternate insect host, Trichoplusia ni. Heterogeneity in one preparation of O. pseudotsugata NPV was observed. The identification procedure is direct and precise. Applications of this procedure include quality control of commercial preparations of viral pesticides and screening for genetic alterations in the viruses.

DNA Restriction Enzymes↗

Isolation of genotypic variants of Autographa californica nuclear polyhedrosis virus.

A nuclear polyhedrosis virus (MNPV) isolated from a lepidopteran (Noctuidae) insect, Autographa californica, was cloned by successive plaque purification using virions containing only one nucleocapsid per envelope as inoculum. The ability to clone the virus by this method was demonstrated by the isolation of nondefective, genotypic variants of the virus with similar but not identical restriction endonuclease fragment patterns. Five distinct variants were identified by genotypic analysis with HindIII, EcoRI, SalI, and Bam HI restriction endonucleases. The characteristic genotype of each variant was maintained upon passage in insect larvae. The isolation of these virus variants demonstrates (i) the heterogeneity of the uncloned virus preparation and (ii) the ability to clone MNPVs by plaque purification of media-derived nonoccluded virions. The A. californica MNPV is being considered for commercial use as a pesticide in the United States, and the cloning of the virus, in view of the heterogeneity detected, may be advisable. The cloning and genotype analyses are also significant with regard to understanding the genetic nature of multiply embedded NPVs (those NPVs containing more than one nucleocapsid per envelope in the occluded form of the virus) and indicate that further genetic analysis of these viruses is possible.

Animals↗

Multiple forms of oviduct progesterone receptors analyzed by ion exchange filtration and gel electrophoresis.

Resolution of the multiple forms of steroid receptors in small samples has been improved by two new techniques: preparative ion exchange filtration and electrophoresis in highly cross-linked polyacrylamide gels of varied concentration. These techniques were used in conjunction with protamine precipitation, gel filtration, and density gradient centrifugation to separate five forms of the progesterone receptor of chick oviduct cytosol. These complexes, numbered I to V in order of elution from agarose gel columns, have been characterized with respect to apparent molecular weight, shape, and relative net charge. Form I, which is eluted in the void volume after gel filtration of cytosol in hypotonic media, is heterodisperse with respect to sedimentation coefficient and electrophoretic mobility (Rf). Form I is converted to form III by KC1. Form II has the highest axial ratio and the highest Rf at pH 10.2. This 4.2S complex can be extracted from DEAE filters, but not from protamine-precipitated cytosol, by 0.3 to 0.5 M KC1. Form III is slightly smaller (3.9S) and less asymmetric than form II. It is relased from DEAE filters and protamine-precipitated cytosol by 0.15 M KC1 and displays increased Rf upon purification. Forms II and III correspond to the B and A components described by W. T. Schrader and B. W. O'Malley ((1972), J. Biol. Chem 247, 51). Form IV may result from the proteolytic cleavage of forms II and/or III. Form V is a globular polypeptide obtained in the presence of certain divalent cations. This complex has been named the "mero-receptor" since it is the smallest part or fragment of the receptor that contains the steroid-binding site.

Animals↗

Fate of mismatched base-pair regions in polyoma heteroduplex DNA during infection of mouse cells.

Heteroduplex DNA has been constructed from two variants of polyoma virus that differ genotypically at four distinct sites. The genotypes of the progeny virus derived from infections of mouse cells with single heteroduplexes have been analyzed to determine how the genotypic markers of the parental heteroduplex segregate. Markers that are separated by a length of DNA greater than 600 nucleotides segregate independently. Segregation was not detected between two markers separated by only approximately 90 nucleotides. We interpret these results on the basis of the correction of mismatched base-pair regions in the heteroduplex before the completion of DNA replication. We suggest that this technique provides valuable information concerning gene conversion in mamalian cells and permits the transfer of genotypic markers from one virus strain to another.

Base Sequence↗

Construction of the genetic map of the polyoma genome.

Seven early mutants, three late mutants, and one plaque morphology mutant of polyoma have been mapped by marker rescue using wild-type restriction endonuclease fragments. The early mutants map between 1.0 and 26.4 units from the Eco RI site, a region previously shown to correspond to the 3'-OH termainal half of "early" RNA (Kamen et al., 1974). The late mutants as well as the plaque morphology mutant map between 26.6 and 45.4 map units, a region previously shown to correspond to the 3'-OH terminal half of "late" RNA (Kamen et al., 1974). Analysis of the genotype of rescued virus demonstrated that the modification of the mutant DNA during marker rescue was limited to the region of the genome covered by the wild-type restriction endonuclease fragment tested.

Chromosome Mapping↗

The design and evaluation of a worksharing system for experimental group living.

Failure to share household chores equitably may be a major cause of the high failure rate of experimental group-living arrangements. A behavioral approach to worksharing based on a point system was implemented in one experimental group and its most important components experimentally evaluated. Experiment I showed that awarding credits produced more work than not awarding credits. Experiment II showed that making credits contingent on the outcome of a detailed inspection produced more work than awarding credits noncontingently. Experiment III demonstrated that awarding rent reductions contingent on credit earnings produced more work than awarding rent reductions noncontingently. Other evaluative data suggest that the resulting living arrangement is cheaper, more effective, and more satisfactory to the residents when compared to the most popular alternative living arrangements.

Journal Article↗

A behavioral technology for producing concept formation in university students.

Three experiments analyzed the effectiveness of a textbook incorporating "concept programming" in producing concept formation in university students. The concept programming portion of each lesson requires students to determine which concept is illustrated by each of 20 short fictional stories about everyday behavioral situations. The stories are selected to illustrate and contrast the concepts of that lesson. Student responses are heavily prompted during the initial stories of each lesson. The first experiment demonstrated that students generalize to entirely novel examples from the examples in the textbook. The second experiment demonstrated that the concept programming portion of the textbook is a critical component in producing generalization. The third experiment demonstrated that the amount of concept formation produced by the concept programmed textbook is greater than that produced by a widely used standard textbook.

Journal Article↗

Steroid-receptor quantitation and characterization by electrophoresis in highly cross-linked polyacrylamide gels.

Conditions for discontinuous polyacrylamide gel electrophoresis have been defined in which progesterone receptors of chick oviduct cytosol and a variety of steroid-binding proteins from other sources are stable and amenable to quantitative analysis. The essential modifications from standard procedures include the use of (1) separation gels in which the cross-linking agent/acrylamide monomer = 15:85, (2) glycerol (10% v/v) in all phases of the Trisglycine-HCl buffer system (pH 10.2 in the separation phase during electrophoresis at 0 degrees), and (3) a layer of a charged reducing agent, thioglycolate, beneath the sample layer. Electrophoresis of untreated oviduct cytosol labeled with [3H]progesterone +/- competing steroids revealed a heterodisperse slow peak and a sharp fast peak. Both peaks displayed the steroid-binding specificity and saturability that are characteristic of intracellular receptors. Recovery of steroid from both the slow and fast components increased linearly with sample load up to 60 mul of cytosol (1.2 mg of protein)/gel (6 mm diameter). The specific progesterone binding detected by this technique was comparable to that detected by charcoal-dextran treatment or ion exchange filtration. Relative electrophoretic mobilities (Rf) of globular protein standards and steroid-protein complexes in cytosol and chick serum were measured in separation gels with total gel concentrations (T) systematically varied from 5 to 15% (w/v). Data were processed by computer programs to obtain weighted linear regressions of log Rf on T (Ferguson plots) and the joint 95% confidence limits of the slopes (-KR) and intercepts of these plots. Molecular radii (R) of the binding components and apparent molecular weights (M) were calculated from the linear correlation of R with KR 1/2 for the standards. The value of M is approximately 158,000 obtained for the cytosol fast component was independent of the length of the separation gel, the presence of a stacking gel or prior exposure of the cytosol to KCl. It was higher than expected from the sedimentation coefficient of 4.2 S in the same pH 10.2 buffer. Electrophoresis in 170-mm separation gels without stacking gels revealed that KCl extracts of protamine-precipitated cytosol contain a different receptor form, of lower net negative charge than the cytosol fast form. The results demonstrate the utility of electrophoresis in highly cross-linked gels of several concentrations to discriminate between various receptor forms and steroid-binding components of serum. This method may lead to overestimates of M for highly asymmetric receptor forms.

Animals↗

Training public-speaking behavior: an experimental analysis and social validation.

The effect of an instructional package on public-speaking behaviors was analyzed in two experiments. The instructional package was designed to teach public-speaking trainees to look at the audience, make gestures, and perform a number of speaking behaviors. The results of Experiment I, with a university student serving as the trainee, showed that the percentage of each category of public-speaking target behavior increased only after the instructional package was introduced for that category. The results of Experiment 2, with three low-income paraprofessional staff members of a neighborhood service center serving as trainees, showed that the percentage of target behaviors increased after the instructional package was introduced for the respective trainee. Audience ratings of public-speaking performance were correlated with direct observations of target responses. All trainees showed marked improvements in audience ratings from pretraining to posttraining. This study demonstrated an effective procedure for training public-speaking behaviors.

Journal Article↗