Biomedical subjects

Thomas E Johnson

Publications and source records attributed to Thomas E Johnson.

63 records · Page 4Linked to original sources

PubMed2002

Heat shock changes the heterogeneity distribution in populations of Caenorhabditis elegans: does it tell us anything about the biological mechanism of stress response?

In this paper we analyze survival data of populations of sterilized nematodes, Caenorhabditis elegans, exposed to heat shocks of different duration at the beginning of their adult lives. There are clear hormesis effects after short exposure to heat and clear debilitation effects after long exposure. Intermediate durations result in a mixture of these two effects. In this latter case, the survival curves for the control and experimental populations intersect. We show that observed effects may be explained by using a model of discrete heterogeneity. According to this model, each population of worms in the experiment is a mixture of subcohorts of frail, normal, and robust individuals; exposure to heat changes the initial proportion of worms in the subcohorts (heterogeneity distribution); and these changes depend on the duration of exposure. In other words, exposure to heat does not influence mortality rates (survival functions) in the subcohorts but does cause individuals to move from one subcohort to another. In a biological interpretation of this finding we hypothesize that, when coping with stress, the organisms of worms use several lines of defense. Switching these lines on and off in response to stress in individual organisms generates the spectrum of observed survival effects at the population level. We discuss possible molecular biological mechanisms of stress response and directions for further research.

Animals↗

PubMed2002

Possible pleiotropic effects of genes specifying sedative/hypnotic sensitivity to ethanol on other alcohol-related traits.

BACKGROUND: Initial sensitivity to ethanol is a predictor of alcohol abuse that has been studied extensively in both human and animal populations. Selection for initial sensitivity to the sedative/hypnotic effects of ethanol resulted in the long-sleep and short-sleep lines of mice. Some of the genes selected in these lines could also specify differential responses in other ethanol-related phenotypes and, perhaps, for other drugs of abuse. We assessed congenic mice carrying a single quantitative trait locus (QTL) from the inbred long-sleep (ILS) or inbred short-sleep (ISS) strain on the reciprocal background for a number of ethanol- and pentobarbital-related phenotypes. METHODS: Each congenic strain was tested for ethanol elimination rates at 4.1 g/kg, ethanol-induced ataxia at 2.0 g/kg, ethanol-induced hypothermia at 4.1 g/kg, and pentobarbital-induced loss of righting reflex (LORR) at 60 mg/kg. Additionally, the ILS.ISS congenics were tested for low-dose ethanol-induced activation (LDA) at five doses ranging from 0.6 to 1.2 g/kg ethanol, and the ISS.ILS congenics were tested for LDA at 1.8 g/kg of ethanol. RESULTS: There was little difference in the ethanol elimination rate between congenics and background strains, although a modest sex effect was found, with the females eliminating ethanol more rapidly than the males. We were unable to replicate previous differences found in LDA for the congenic on the ISS background, because none of the congenics differed from controls for LDA. congenics showed a differential effect of pentobarbital-induced LORR in the expected directions. The congenics on the ISS background showed more ethanol-induced ataxia than the ISS controls. Additionally, the hypothermic response seems affected by and and maybe others. CONCLUSIONS: At least two regions carrying a QTL specifying sensitivity to high doses of ethanol cospecify altered sensitivity in other measures of alcohol action. Specifically, these QTLs clearly affect ethanol-induced hypothermia and pentobarbital-induced LORR and possibly ethanol-induced ataxia.

Alcoholism↗

PubMed2002

Fine mapping of polymorphic alcohol-related quantitative trait loci candidate genes using interval-specific congenic recombinant mice.

BACKGROUND: The inbred long-sleep (ILS) and inbred short-sleep (ISS) strains of mice are widely studied as a model of initial sensitivity to alcohol. Recently, a large comparative DNA sequencing study of candidate genes located within the four Lore quantitative trait loci (QTLs) associated with the ethanol-induced loss of righting reflex in ILS and ISS mice has identified eight genes that contain coding region differences corresponding to amino acid changes. Here, recently developed interval-specific congenic recombinant mice (ISCRs) have been used to map these genes in relationship to newly narrowed QTL regions. METHODS: Regions of candidate genes containing DNA differences corresponding to previously identified amino acid changes between ISS and ILS mice were amplified from either genomic DNA or complementary DNA from ISCR mice using polymerase chain reaction. The products were purified and directly sequenced to determine the genotypes for each polymorphism. On the basis of these genotypic data, each candidate gene was determined to be located either within or outside of recently narrowed Lore QTL intervals. RESULTS: Of these eight candidates with protein-coding differences, five are now excluded from their respective Lore intervals. The other three (Znf142, Ptprn, and Znf133) have been localized to the narrowed QTL intervals. CONCLUSIONS: These three central nervous system genes (Znf142, Ptprn, and Znf133) represent promising candidates for involvement in the differential sensitivity to alcohol exhibited between ILS and ISS mice. This study also demonstrates how the combination of high-throughput comparative gene sequencing and concomitant genetic fine mapping of QTL regions with ISCRs can be an effective tool for accelerating the process of moving from QTL to gene.

Alcoholism↗

PubMed2002

Genetic dissection of quantitative trait loci specifying sedative/hypnotic sensitivity to ethanol: mapping with interval-specific congenic recombinant lines.

BACKGROUND: Linkage studies alone do not produce sufficient resolution to narrow the location of a quantitative trait locus (QTL) to a small-enough chromosomal region for gene identification. One solution to this problem is to use interval-specific congenic recombinant (ISCR) lines to narrow the chromosomal interval known to contain the QTL. In previous work, we mapped four QTLs for differential ethanol sensitivity in the inbred long-sleep (ILS) and inbred short-sleep (ISS) strains and generated reciprocal congenic strains in which each full QTL interval from ILS was bred onto the ISS background and vice versa. METHODS: ISCR lines were derived by identifying mice carrying recombination events in the congenic interval during backcrossing of the ISS.ILS.Lore congenics to ISS. Recombinant mice were backcrossed to ISS, and progeny carrying the ISCR chromosome were identified and tested to determine whether the ISCR region carried the donor Lore QTL. RESULTS: We developed multiple ISCR lines for each Lore QTL, in which the QTL interval was broken into a number of smaller intervals. For all four QTLs, we reduced the size of the interval, in one case to 3.7 cM. CONCLUSIONS: Use of ISCR lines can narrow each Lore candidate region to a few centimorgans. Such an interval size is conducive to brute-force approaches to identify candidate genes, entailing bioinformatics, gene expression, and DNA sequencing strategies.

Alcohol Drinking↗

PubMed2002

Identification of a novel cis-regulatory element involved in the heat shock response in Caenorhabditis elegans using microarray gene expression and computational methods.

We report here the identification of a previously unknown transcription regulatory element for heat shock (HS) genes in Caenorhabditis elegans. We monitored the expression pattern of 11,917 genes from C. elegans to determine the genes that were up-regulated on HS. Twenty eight genes were observed to be consistently up-regulated in several different repetitions of the experiments. We analyzed the upstream regions of these genes using computational DNA pattern recognition methods. Two potential cis-regulatory motifs were identified in this way. One of these motifs (TTCTAGAA) was the DNA binding motif for the heat shock factor (HSF), whereas the other (GGGTGTC) was previously unreported in the literature. We determined the significance of these motifs for the HS genes using different statistical tests and parameters. Comparative sequence analysis of orthologous HS genes from C. elegans and Caenorhabditis briggsae indicated that the identified DNA regulatory motifs are conserved across related species. The role of the identified DNA sites in regulation of HS genes was tested by in vitro mutagenesis of a green fluorescent protein (GFP) reporter transgene driven by the C. elegans hsp-16-2 promoter. DNA sites corresponding to both motifs are shown to play a significant role in up-regulation of the hsp-16-2 gene on HS. This is one of the rare instances in which a novel regulatory element, identified using computational methods, is shown to be biologically active. The contributions of individual sites toward induction of transcription on HS are nonadditive, which indicates interaction and cross-talk between the sites, possibly through the transcription factors (TFs) binding to these sites.

Animals↗

PubMed2002

Evaluation of a lateral-flow immunoassay for use in monitoring passive transfer of immunoglobulins in calves.

OBJECTIVE: To determine the predictive ability of a commercially available lateral-flow immunoassay used for determining passive transfer of immunoglobulins in calves. ANIMALS: 204 male Holstein calves ranging from 4 to 8 days old. PROCEDURE: Serum samples were obtained from each calf. Results of refractometry, zinc sulfate turbidity technique, and the lateral-flow immunoassay were determined. Sensitivity, specificity, accuracy, and predictive ability were calculated on the basis of IgG concentrations determined by turbidimetric immunoassay (TIA). RESULTS: Mean IgG concentration in the study was 10.9 mg/ml as determined by TIA. Rate of failure of passive transfer in this study population was 56%. Associations between the values for the refractometry and zinc sulfate turbidity techniques were established by regression analysis. Accuracy for the lateral-flow immunoassay, refractometry, and zinc sulfate turbidity methods was 95, 80, and 73%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The lateral-flow immunoassay was better at determining the status of passive transfer of immunoglobulins, compared with the refractometry or zinc sulfate turbidity methods. The ability of the lateral-flow immunoassay to provide accurate results should enable clinicians to make immediate management or intervention decisions.

Animals↗

PubMed2002

Corneal injury threshold in rabbits for the 1540 nm infrared laser.

BACKGROUND: In the 40 yr since lasers were invented, they have become commonplace in military operations, and while their utility in this setting is undeniable, they also represent a potential hazard for those in contact with them. This threat must be recognized, information must be gathered to understand this injury potential, and the necessary measures must be taken to properly protect those who will work, train, and fight with these systems. The exact mechanisms of laser/tissue interaction at 1540 nm are not well understood. Previous studies and textbooks show remarkable disparity in reporting where 1540 nm laser energy is deposited and the quantity of energy required to cause tissue damage. Rabbit cornea is very similar histologically to that of humans with the exception that it lacks Bowman's membrane. This model has been recommended as a reasonable approximation by past researchers and avoids the use of valuable non-human, primate research animals. METHODS: A rabbit model was used to demonstrate the ability of the 1540 nm laser to produce corneal injuries. Various energies were applied to find the threshold at which injury is consistently produced. Observations included the appearance of the injury in the rabbit cornea. All rabbits were between 5 and 6 kg. RESULTS: Corneal injury was consistent at energies above 56 J x cm(-2). Injuries involved the deeper corneal stroma rather than only the epithelial layer, thus raising concern for permanent visual disability in those affected. The gross appearance of these injuries was white opaque areas easily seen within the corneal stroma. CONCLUSIONS: Data shows conclusively that the 1540 nm laser causes significant corneal damage at reproducible energy levels. Further research is clearly necessary to advance our understanding of the role of Bowman's membrane, the healing properties of the injured cornea, and the epidemiology of laser injury.

Animals↗

PubMed2002

Study of corneal lesions induced by 1,318-nm laser radiation pulses in Dutch belted rabbits (Oryctolagus cuniculus).

BACKGROUND AND PURPOSE: Use of high-energy near-infrared lasers is becoming more prevalent in today's industries, such as technology, medicine, and military operations. Despite wide-range use of these lasers, threshold, median effective dose (ED50), and the mechanism of laser-tissue interaction are not well defined at the 1,318-nm wavelength for human corneal exposures. The goals of the study reported here were to establish the ED50 for single-pulse, 1,318-nm laser exposures on the Dutch Belted rabbit cornea and to characterize microscopic changes. Results of this study were then compared with those of previous corneal studies. METHODS: A neodymium:yttrium aluminum garnet (Nd:YAG) laser was used to deliver single 1,318-nm wavelength pulses to the corneas of 10 female Dutch Belted rabbits (Oryctolagus cuniculus). Single pulses of 0.5-ms duration and radiant energy ranging from 116 to 2,250 J/cm2 irradiated the exposure sites. Sites were clinically evaluated for presence of a lesion at one hour and 24-h after exposure. Results of the 24-h evaluation were used to determine the (ED50). Corneas were subsequently collected at the 24-h endpoint for microscopic evaluation. RESULTS: The ED50 for 1,318-nm exposures to the rabbit cornea was determined to be 382 J/cm2, as measured at the 1/e2 (0.865 times that of the peak power per unit area). At each exposure site, there was a small (< 1 mm in diameter), white, circular, well demarcated corneal lesion characterized histologically by a band of stromal coagulative necrosis and endothelial necrosis, with sparing of the anterior epithelium. In addition, there appeared some potential for damage to Descemet's membrane at the highest energy level tested. CONCLUSIONS: Findings indicate that the rabbit corneais subject to injury at the 1,318-nm wavelength with the established ED50.

Animals↗

PubMedDate not supplied

Advantages and disadvantages of Caenorhabditis elegans for aging research.

The nematode Caenorhabditis elegans has been the organism of choice for most aging research, especially genetic approaches to aging. More than 70 longevity genes have been identified, with more to come, and these genes have been the subjects of intense study. I identify the major reasons for this and discuss limitations of this organism for future progress in research on aging.

Aging↗