Keratin polypeptide expression in mouse epidermis and cultured epidermal cells

Adult mouse epidermis contains up to 11 distinct keratin polypeptides, as resolved by two-dimensional gel electrophoresis. These include both basic (Type II; 67-, 65-, 63-, 62-, and 60-kDa) and acidic (Type I; 61- to 59-, 54-, 52-, 49-, and 48-kDa) keratins that exhibit multiple isoelectric forms. Several, but not all, of these keratins, identified by immunoblotting, were found to be actively synthesized in the skin when assayed in short-term pulse-labeling experiments. When compared to the adult, newborn mouse epidermis expresses fewer keratin subunits. However, greater amounts of keratins associated with differentiated suprabasal cells and stratum corneum, which is more pronounced morphologically in the newborn, were identified. We also observed strain-specific differences in the expression of a Type I acidic keratin. This 61-kDa (pI, approx. 5.3) keratin was produced exclusively by the CF-1 mouse and, based on peptide mapping, appeared to be related to the acidic 59-kDa keratin that was identified in this strain as well as all other mouse strains. The 61-kDa keratin was not expressed in vitamin A-deficient animals, suggesting that its appearance may be related to a retinoid-dependent posttranslational modification. In comparison to keratin expression in vivo, primary mouse keratinocyte monolayer cultures maintained in low Ca2+ (less than 0.08 mM) did not express the terminal differentiation keratins of 67-kDa (basic) or 59-kDa (acidic), although enhanced synthesis of the 60-kDa (basic) and the 52-kDa and 59-kDa (acidic) keratins associated with proliferation were observed. In addition, a subpopulation of nonadherent cells was continuously produced by the primary keratinocyte cultures that expressed the 67-kDa (basic) keratin specific for terminal differentiation. When the keratinocyte cultures were induced to terminally differentiate with Ca2+, the overall pattern of keratin expression was not changed significantly. Taken together, these results provide further evidence for the variable nature of keratin expression in mouse epidermal keratinocytes under different growth conditions.     

Surface EMG recordings and pain-related disorders: A diagnostic framework

Surface electromyography provides a very valuable set of information when used diagnostically with pain-related disorders. Unfortunately, most researchers and clinicians limit their investigation to only one level of diagnostic information available concerning the neuromuscular system. This article develops and encourages the clinician/researcher to consider three levels of diagnostic information: emotional, organ-related, and postural aspects. The theoretical background and diagnostic procedures used to investigate each is presented. Differential treatment considerations, given differential diagnostic findings, are discussed.     

Persistence of community structure: what happens when you change taxonomic scale?

It is common in community ecology to use the species as the taxonomic category of interest, yet in rich tropical assemblages containing guilds of very similar species this may not be appropriate. Such assemblages may be organized at the level of guilds rather than at the finer species level. In a ten-year study of assemblages of fish at One Tree Reef, Great Barrier Reef, we found species composition and the number of fish on a given lagoonal patch reef vary greatly across time (Sale and Douglas 1984; Sale et al. in preparation). The mean average proportional similarity of a reef's assemblage to itself at different times (censuses) is usually low at a value of around 0.5. This apparent variability may be ecologically irrelevant noise if organization is at the higher guild level. We have recast our database at the guild level to test this possibility. Thirteen guilds were defined by the diets, foraging habitats and times of the individual species comprising them. Similarity of an assemblage to itself at successive censuses was re-calculated using the number of individuals in each guild instead of the numbers in each species. This analysis yielded significantly higher levels of similarity (P<0.01) among censuses. To test whether this increase in similarity was due solely to the smaller number of categories used to calculate the similarity indices, 5 sets of randomly generated guilds were constructed using a Monte Carlo approach. No significant difference (P>0.05) was found between the average similarity among censuses when assemblages were classified into these null guilds and when they were classified according to the real guilds. These results indicate that shifting to the larger taxonomic scale of guilds does not reveal a significantly more persistent assemblage structure than that revealed when analysis is at the smaller scale of species. There is no evidence of an underlying organization of these assemblages at the guild level.     

Alu-PCR: characterization of a chromosome 6-specific hybrid mapping panel and cloning of chromosome-specific markers.

The Alu-polymerase chain reaction (Alu-PCR) was applied to selectively amplify DNA sequences from human chromosome 6 using a single primer (A1) directed to the human Alu consensus sequence. A specific amplification pattern was demonstrated for a panel of eight somatic cell hybrids containing different portions of chromosome 6. This PCR pattern permits the identification of submicroscopic DNA alterations and can be utilized as a reference for additional chromosome 6-specific hybrids. To obtain new chromosome 6-specific markers we established two libraries from PCR-amplified sequences using two somatic cell hybrids (MCH381.2D and 640-5A). Out of a total of 109 clones that were found to be chromosome 6 specific, 13 clones were regionally assigned. We also included a procedure that allows the isolation of chromosome 6-specific markers from hybrids that contain human chromosomes other than 6. Our results will contribute to the molecular characterization of chromosome 6 by fostering characterization of somatic cell hybrids and by the generation of new regionally assigned DNA markers.     

Biologic markers in ethylene oxide-exposed workers and controls

Ethylene oxide (EtO) is an alkylating agent and a model direct-acting mutagen and carcinogen. This study has evaluated a panel of biologic markers including EtO-hemoglobin adducts (EtO-Hb), sister-chromatid exchanges (SCEs), micronuclei, chromosomal aberrations (CAs), DNA single-strand breaks (SSB) and an index of DNA repair (ratio of UDS to NA-AAF-DNA binding) in the peripheral blood cells of 34 workers at a sterilization unit of a large university hospital and 23 controls working in the university library. Comprehensive environmental histories were obtained on each subject including detailed occupational and smoking histories. Industrial hygiene data obtained prior to the study and personal monitoring during the 8 years preceding the study showed that workers were subject to low-level exposure near or below the current Occupational Safety and Health Administration (OSHA) standard of 1 ppm (TWA). Personal monitoring data obtained during 2 weeks prior to blood sampling were uniformly less than 0.3 ppm (TWA). After adjusting for smoking, EtO workplace exposure was significantly (p less than 0.001) associated with EtO-Hb (a carcinogen-protein adduct) and 2 measures of SCEs [the average number of SCEs/cell (SCE50) and the number of high frequency cells (SCEHFC)]. There was an apparent suppression of DNA repair capacity in EtO-exposed individuals as measured by the DNA repair index; i.e., the ratio of unscheduled DNA synthesis (UDS) and NA-AAF-DNA binding (p less than 0.01). No association of DNA repair index with smoking was found. Another important finding of this study is the highly significant correlation between EtO-Hb adduct levels and SCEHFC (p less than 0.01) and SCEs (p less than 0.02) which provides evidence of a direct link between a marker of biologically effective dose and markers of genotoxic response. In contrast, micronuclei, CAs and SSBs were not significantly elevated in the workers. The activity of the u-isoenzyme of glutathione-S-transferase (GT) was measured as a possible genetic marker of susceptibility and a modulator of biomarker formation. However, possibly because of confounding by age, no significant relationships were found between GT and any of the exposure-related markers by ANOVA or among other independent variables by regression. This study demonstrates significant effects of low-level EtO exposure, independent of smoking history, near or below 1 ppm on multiple biomarkers and suggests that the current OSHA standard may not be adequately protective. Previously described effects of smoking on EtO-Hb adducts, SCEs and SCEHFC were also seen in this study.(ABSTRACT TRUNCATED AT 400 WORDS)     

Environmental effects on variation and covariation in reproductive traits of Western Bluebirds

Summary Environmental conditions can influence the expression and correlations of phenotypic traits. I studied phenotypic plasticity in reproductive traits of Western Bluebirds breeding in northern Arizona. Data collected over 4 years on two contrasting habitats identified significant spatial and temporal variation in bluebird reproduction. Clutch size was similar over different environmental conditions whereas timing of clutch initiation, percent fledging success, and frequency of second nest attempts were flexible. Correlations between traits varied widely—often changing sign—among samples from different years or habitats. Correlations of traits with reproductive success were also dependent on environmental conditions. Variation in traits reflected behavioral responses by nesting adults to differences in time for breeding and feeding conditions. Density of trees differed between habitats and had opposing effects on these environmental variables; breeding seasons were generally longer, but feeding rates to nestlings were lower on the more open habitat. Late Spring snows delayed reproduction and increased the importance of limited time for breeding; feeding conditions were more influential following a dry Spring. This and other studies illustrate that data on phenotypic plasticity are important when evaluating the ecological and evolutionary forces underlying life histories.     

A method for determining overall protein fold from NMR distance restraints

Summary We describe a simple method for determining the overall fold of a polypeptide chain from NOE-derived distance restraints. The method uses a reduced representation consisting of two particles per residue, and a force field containing pseudo-bond and pseudo-angle terms, an electrostatic term, but no van der Waals or hard shell repulsive terms. The method is fast and robust, requiring relatively few distance restraints to approximate the correct fold, and the correct mirror image is readily determined. The method is easily implemented using commercially available molecular modeling software.     

The sinking ark: pollution and the worldwide loss of biodiversity

Humans are making increasing demands on natural ecosystems. One recent study has concluded that our species is consuming or diverting some 40% of the net photosynthetic productivity of our planet. Many habitats are being converted to simpler systems which provide more harvestable goods to people. As a result, genetic diversity, species and whole ecosystems are disappearing; some scientists suggest that as many as 25% of the world's species could be lost in the next several decades.The sinking ark is usually characterized in terms of pollution, habitat loss, poaching, introduced species and illegal trade in wildlife products, but these are symptoms rather than causes. At a more fundamental level, many of the same factors which have enabled pollution to become such a problem have also been responsible for the loss in biodiversity; the most important factor is that the effects of pollutants on biodiversity have been considered an externality, an unintended side effect of industrial activity which brought measurable benefits to people. Development activities which have depleted biodiversity have proven profitable only because the real costs have been hidden.Keeping the ark afloat will require the Five-I Approach: investigation (learning how natural systems function); information (ensuring that the facts are available to inform decisions); incentives (using economic tools to help conserve biodiversity); integration (promoting a cross-sectoral approach to conserving biodiversity); and international support (building productive collaboration for conserving biodiversity).