Rapid morphological change in black rats (Rattus rattus) after an island introduction.

Rapid morphological change has been shown in rodent populations on islands, including endemic deer mice (Peromyscus maniculatus subspp.) on the California Channel Islands. Surprisingly, most of these changes were towards a smaller size. Black rats were introduced to Anacapa Island in the mid-1800s (probably in 1853) and eradicated in 2001-2002. To assess possible changes in these rats since their introduction, eleven cranial and four standard external measurements were taken from 59 Rattus rattus specimens collected from 1940-2000. All rat cranial traits changed 3.06-10.43% (724-2567 d, 0.06-0.42 h), and all became larger. When considered in haldanes, these changes are among the fastest on record in any organism, and far exceed changes found in other island rodents. These changes were confirmed by MANOVA (Wilk's λ < 0.0005, F d.f.15 = 2974.386, P < 0.0005), and all 11 cranial traits significantly fit linear regressions. We speculate that concurrent changes in mice may have been due in part to competition with and/or predation by rats. Future research might evaluate whether the vector of mouse evolution on Anacapa is again changing after rat eradication.

The impact of nature experience on willingness to support conservation.

We hypothesized that willingness to financially support conservation depends on one's experience with nature. In order to test this hypothesis, we used a novel time-lagged correlation analysis to look at times series data concerning nature participation, and evaluate its relationship with future conservation support (measured as contributions to conservation NGOs). Our results suggest that the type and timing of nature experience may determine future conservation investment. Time spent hiking or backpacking is correlated with increased conservation contributions 11-12 years later. On the other hand, contributions are negatively correlated with past time spent on activities such as public lands visitation or fishing. Our results suggest that each hiker or backpacker translates to $200-$300 annually in future NGO contributions. We project that the recent decline in popularity of hiking and backpacking will negatively impact conservation NGO contributions from approximately 2010-2011 through at least 2018.

Recent and widespread rapid morphological change in rodents.

In general, rapid morphological change in mammals has been infrequently documented. Examples that do exist are almost exclusively of rodents on islands. Such changes are usually attributed to selective release or founder events related to restricted gene flow in island settings. Here we document rapid morphological changes in rodents in 20 of 28 museum series collected on four continents, including 15 of 23 mainland sites. Approximately 17,000 measurements were taken of 1302 rodents. Trends included both increases and decreases in the 15 morphological traits measured, but slightly more trends were towards larger size. Generalized linear models indicated that changes in several of the individual morphological traits were associated with changes in human population density, current temperature gradients, and/or trends in temperature and precipitation. When we restricted these analyses to samples taken in the US (where data on human population trends were presumed to be more accurate), we found changes in two additional traits to be positively correlated with changes in human population density. Principle component analysis revealed general trends in cranial and external size, but these general trends were uncorrelated with climate or human population density. Our results indicate that over the last 100+ years, rapid morphological change in rodents has occurred quite frequently, and that these changes have taken place on the mainland as well as on islands. Our results also suggest that these changes may be driven, at least in part, by human population growth and climate change.

Evidence for a fundamental and pervasive shift away from nature-based recreation.

After 50 years of steady increase, per capita visits to U.S. National Parks have declined since 1987. To evaluate whether we are seeing a fundamental shift away from people's interest in nature, we tested for similar longitudinal declines in 16 time series representing four classes of nature participation variables: (i) visitation to various types of public lands in the U.S. and National Parks in Japan and Spain, (ii) number of various types of U.S. game licenses issued, (iii) indicators of time spent camping, and (iv) indicators of time spent backpacking or hiking. The four variables with the greatest per capita participation were visits to Japanese National Parks, U.S. State Parks, U.S. National Parks, and U.S. National Forests, with an average individual participating 0.74-2.75 times per year. All four time series are in downtrends, with linear regressions showing ongoing losses of -1.0% to -3.1% per year. The longest and most complete time series tested suggest that typical declines in per capita nature recreation began between 1981 and 1991, are proceeding at rates of -1.0% to -1.3% per year, and total to date -18% to -25%. Spearman correlation analyses were performed on untransformed time series and on transformed percentage year-to-year changes. Results showed very highly significant correlations between many of the highest per capita participation variables in both untransformed and in difference models, further corroborating the general downtrend in nature recreation. In conclusion, all major lines of evidence point to an ongoing and fundamental shift away from nature-based recreation.

Rapid morphological and genetic change in Chicago-area Peromyscus.

We report rapid change of morphology and mitochondrial genes in white-footed mice (Peromyscus leucopus) in the Chicago (Illinois, USA) region. We sequenced mitochondrial DNA COX2 from 55 museum skins of white-footed mice caught in the Chicago area since 1855 and from 44 mice recently trapped in the same locations. We found consistent directional genotype replacement at five separate collection locations. We later focused on a single one of these locations (Volo Bog State Natural Area) and sequenced mitochondrial D-loop control region from 58 museum skins of mice collected in 1903-1976 and 32 mice recently trapped there. We found complete and more recent replacement of D-loop haplotypes, apparently occurring between 1976 and 2001. We tested whether these genetic changes were mirrored by changes in morphology by comparing 15 external and cranial traits. We found no significant morphological differences between mice collected in 1903-1976; however, mice collected in 2001-2003 showed 9 of 15 measurements to be significantly changed relative to the earlier samples. Recent mice were longer in total length, with broader, longer noses, and longer but shallower skulls(1). Discriminant function analysis allowed for 100% correct classification using these traits. Principal components analysis shows variance over time is well distributed across both external and cranial measures. The sequential replacements of haplotypes and the rapid change of morphology can best be explained by replacement of the regional population with immigrants from genetically distinct neighbouring populations, likely facilitated by the large environmental changes occurring over the time period. Replacement with genotypes from external populations may be a common mechanism of evolution of newly adaptive local forms in an increasingly human-impacted world.

Is love of nature in the US becoming love of electronic media? 16-year downtrend in national park visits explained by watching movies, playing video games, internet use, and oil prices.

After 50 years of steady increase, per capita visits to US national parks have declined since 1988. This decline, coincident with the rise in electronic entertainment media, may represent a shift in recreation choices with broader implications for the value placed on biodiversity conservation and environmentally responsible behavior. We compared the decline in per capita visits with a set of indicators representing alternate recreation choices and constraints. The Spearman correlation analyses found this decline in NPV to be significantly negatively correlated with several electronic entertainment indicators: hours of television, (rs=-0.743, P<0.001), video games (rs=-0.773, P<0.001), home movies (rs=-0.788, P<0.001), theatre attendance (rs=-0.587, P<0.025) and internet use (rs=-0.783, P<0.001). There were also significant negative correlations with oil prices (rs=-0.547, P<0.025), foreign travel (rs=-0.452, P<0.05), and Appalachian Trail hikers (rs=-0.785, P<0.001). Income was significantly positively correlated with foreign travel (rs=0.621, P<0.005) but negatively correlated with national park visits (rs=-0.697, P<0.005). There was no significant correlation of mean number of vacation days, indicating available vacation time is probably not a factor. Federal funding actually increased during this period, and so was rejected as a probable factor. Park capacity was rejected as limiting since both total overnight stays and visits at the seven most popular parks rose well into the mid-1990s. Aging of baby boomers was also rejected as they are only now reaching retirement age, and thus during the period of visitation decline were still of prime family vacation age. Multiple linear regression of four of the entertainment media variables as well as oil prices explains 97.5% of this recent decline (r=0.975, multiple r2=0.950, adjusted multiple r2=0.925, SE=0.015, F=37.800, P<0.0001). We may be seeing evidence of a fundamental shift away from people's appreciation of nature (biophilia, Wilson 1984) to 'videophilia,' which we here define as "the new human tendency to focus on sedentary activities involving electronic media." Such a shift would not bode well for the future of biodiversity conservation.

RAPID MORPHOLOGICAL CHANGE IN CHANNEL ISLAND DEER MICE.

Deer mice, Peromyscus maniculatus, collected over 90 years from three California Channel Islands, were examined for evidence of morphological change. Rapid morphological change has occurred in the endemic subspecies from Santa Barbara (P. m. elusus), Anacapa (P. m. anacapae), and Santa Cruz Island (P. m. santacruzae). Data were divided into two temporal classes, 1897-1941 and 1955-1988. Of the 16 morphological characters measured, between five and 10 measures changed significantly (P ⩽ 0.05) with temporal class in each subspecies, and multivariate test statistics were significant (P ⩽ 0.05) for all three subspecies. For each subspecies, depth of braincase, total length, tail length, and hind foot length became smaller over time, except depth of braincase, which became larger in P. m. elusus. The rates of change dramatically exceed those estimated from paleontological records and are even higher than those reported in some experimental selection studies. Temporal change in two characters exceeds differentiation between subspecies. Although changing, each subspecies remained well differentiated, and incorporation of temporal change allowed correct classification of most specimens. Unlike nearly all previous reports of rapid evolution, the changes in these populations were not associated with a founder events or recent introductions. This study demonstrates that rapid phenotypic change can occur in long-established natural populations and temporal stability of morphological characters in such populations, even over short evolutionary time periods, cannot be assumed.

Receptor recognition sites reside in both lobes of human serum transferrin.

The binding of iron by transferrin leads to a significant conformational change in each lobe of the protein. Numerous studies have shown that the transferrin receptor discriminates between iron-saturated and iron-free transferrin and that it modulates the release of iron. Given these observations, it seems likely that there is contact between each lobe of transferrin and the receptor. This is the case with chicken transferrin, in which it has been demonstrated unambiguously that both lobes are required for binding and iron donation to occur [Brown-Mason and Woodworth (1984) J. Biol. Chem. 259, 1866-1873]. Further support to this contention is added by the ability of both N- and C-domain-specific monoclonal antibodies to block the binding of a solution containing both lobes [Mason, Brown and Church (1987) J. Biol. Chem. 262, 9011-9015]. In the present study a similar conclusion is reached for the binding of human serum transferrin to the transferrin receptor. With the use of recombinant N- and C-lobes of human transferrin produced in a mammalian expression system, we show that both lobes are required to achieve full binding. (Production of recombinant C-lobe in the baby hamster kidney cell system is reported here for the first time.) Each lobe is able to donate iron to transferrin receptors on HeLa S3 cells in the presence of the contralateral lobe. The results are not identical with the chicken system, because the C-lobe alone shows a limited ability to bind to receptors and to donate iron. Further complications arise from the relatively weak re-association between the two lobes of human transferrin compared with the re-association of the ovotransferrin lobes. However, domain-specific monoclonal antibodies to either lobe block the binding of N- and C-lobe mixtures in the human system, thus substantiating the need for both.

Association of the two lobes of ovotransferrin is a prerequisite for receptor recognition. Studies with recombinant ovotransferrins.

Different recombinant N-lobes of chicken ovotransferrin (oTF/2N) have been isolated from the tissue-culture medium of baby hamster kidney cells transfected with the plasmid pNUT containing the relevant DNA coding sequence. Levels of up to 40, 55 and 30 mg/1 oTF/2N were obtained for constructs defining residues 1-319, 1-332 and 1-337-(Ala)3 respectively. In addition, a full-length non-glycosylated oTF was expressed at a maximum of 80 mg/1 and a foreshortened oTF consisting of residues 1-682 was expressed at a level of 95 mg/l. These preparations were then used to produce, proteolytically, two different C-lobes (oTF/2C) comprising residues 342-686 and 342-682. The purified recombinant N-lobes (oTF/2N) are similar to the proteolytically derived half-molecule with regard to immunoreactivity and spectral properties; they show some interesting differences in thermal stability. A sequence analysis of the cDNA revealed six changes at the nucleotide level that led to six differences in the amino acid sequence compared with that reported by Jeltsch and Chambon [(1982) Eur. J. Biochem. 122, 291-295]. Electrospray mass spectrometry gives results consistent with these six changes. Interaction between the various N- and C-lobes was measured by titration calorimetry. Studies show that only those lobes that associate in solution are able to bind to the receptors on chick embryo red blood cells. These findings do not support a previous report by Oratore et al.

Production and isolation of the recombinant N-lobe of human serum transferrin from the methylotrophic yeast Pichia pastoris.

The N-lobe of human serum transferrin has been expressed in the methylotrophic yeast Pichia pastoris by placing the hTF/2N cDNA under the control of the methanol-inducible alcohol oxidase promoter. Following induction with methanol, the N-lobe was efficiently secreted into a basal salt medium in shake flasks at a level of 150-240 mg/liter. As judged by mobility on SDS-PAGE, immunoreactivity with two domain-specific monoclonal antibodies, and both thermal stability and spectral properties (indictative of correct folding and ability to bind iron), the recombinant N-lobe produced by the yeast cells appears to be identical to that produced in a mammalian expression system. Electrospray-mass spectrometry and a third domain specific antibody, however, show that approximately 80% of the protein from the yeast cells contains one or two hexose residues.

Posttranslational processing of recombinant human interferon-gamma in animal expression systems.

We have characterized the heterogeneity of recombinant human interferon-gamma (IFN-gamma) produced by three expression systems: Chinese hamster ovary cells, the mammary gland of transgenic mice, and baculovirus-infected Spodopera frugiperda (Sf9) insect cells. Analyses of whole IFN-gamma proteins by electrospray ionization-mass spectrometry (ESI-MS) from each recombinant source revealed heterogeneous populations of IFN-gamma molecules resulting from variations in N-glycosylation and C-terminal polypeptide cleavages. A series of more specific analyses assisted interpretation of maximum entropy deconvoluted ESI-mass spectra of whole IFN-gamma proteins; MALDI-MS analyses of released, desialylated N-glycans and of deglycosylated IFN-gamma polypeptides were combined with analyses of 2-aminobenzamide labeled sialylated N-glycans by cation-exchange high-performance liquid chromatography. These analyses enabled identification of specific polypeptide cleavage sites and characterization of associated N-glycans. Production of recombinant IFN-gamma in the mammalian expression systems yielded polypeptides C-terminally truncated at dibasic amino acid sites. Mammalian cell derived IFN-gamma molecules displayed oligosaccharides with monosaccharide compositions equivalent to complex, sialylated, or high-mannose type N-glycans. In contrast, IFN-gamma derived from baculovirus-infected Sf9 insect cells was truncated further toward the C-terminus and was associated with neutral (nonsialylated) N-glycans. These data demonstrate the profound influence of host cell type on posttranslational processing of recombinant proteins produced in eukaryotic systems.

Mass spectrometric composition and molecular mass of Lumbricus terrestris hemoglobin: a refined model of its quaternary structure.

Sedimentation equilibrium measurements and scanning transmission electron microscopy (STEM) mass mapping of the extracellular, hexagonal bilayer hemoglobin (HBL Hb) of the earthworm Lumbricus terrestris provided masses of 3.41 to 3.66 MDa and 3.56 (+/- 0.13) MDa, respectively. The Hb also contains 57.2 (+/- 6.0) moles of tightly bound Ca per mole of protein. The Hb and its subunits obtained by dissociation, in native, dehemed and reduced carbamidomethylated forms, were subjected to electrospray ionization mass spectroscopy (ESI-MS). Maximum entropy deconvolution identified three groups of peaks, at approximately 16 kDa, 24 to 32 kDa and approximately 53 kDa corresponding to the monomer subunit M (globin chain d), four linker subunits and the disulfide-bonded trimer T (globin chains a + b + c). Subunit M consisted of three components, d1 (15, 992.4), d2 (15, 978.0) and d3 (15, 962.1) (+/- 1.0 Da), with relative intensities 1.0:5:0.3, respectively. Subunit T consisted of four major components, T1 (52, 922.6), T2 (52, 760.0), T3 (52, 598.5) and T4 (52, 435.4) (+/- 4.0 Da), with relative intensities 0.6:1.0:0.2:0.7, respectively. ESI-MS of carbamidomethylated T, demonstrated that, unlike chains b (16, 254.4) and c (17, 289.2), chain a exists as a series of four, hexose-connected, glycosylated isoforms, a1 to a4 (19, 389.9, 19, 227.4, 19, 065.3 and 18, 902.9) (+/- 1.0 Da). The mass differences between the deglycosylated chain a (17, 524.0) and a1 to a4 correspond to glycan side-chains (GlcNAc)2 (Man)n (n = 6 to 9). Four groups of peaks were observed in the 24 to 32 kDa region. Linkers L1a (27, 540.8) and L1b (27, 702.4) (+/- 2.0 Da) are isoforms of L1 (25, 837.5 in N-deglycosylated Hb) with glycan side-chains (GlcNAc)2 (Man)n (n = 8,9). Linkers L2 (32, 104.3 (+/- 5.0) Da) and L3 (24, 912.9 (+/- 2.0) Da) occur as single species. Linkers L4a to L4c (24, 019.0, 24, 102.3 and 24, 169.9) (+/- 2.0 Da) with relative intensities 1.0:0.8:0.8, have not been identified previously. From ESI-MS relative intensities, L1:L2:L3:L4 = 0.6:0.4:1.0:0.5 and globin linker = 0.78:0.22. HPLC of Lumbricus Hb provided a globin linker = 0.73:0.27 (+/- 0.02) and a heme content of 2.52 (+/- 0.14) wt%. A model is proposed for the HBL structure, wherein 12 213.4 kDa dodecamers (144 globin chains, 2561 kDa) decorate a hexagonal framework of 36 linker chains (12L1 + 6L2 + 12L3 + 6L4) to provide a total mass of 3.531 MDa, each dodecamer being in contact with three linker subunits.

Characterization of sheep hemopexin glycovariants.

The hemopexin phenotype HpxB1 isolated from sheep serum, yields three major bands when subjected to starch gel and/or polyacrylamide gel electrophoresis which are here designated as subcomponents HpxB1-I, HpxB1-II and HpxB1-III. Electrospray mass spectrometric analysis of samples of the isolated subcomponents prepared by ion exchange chromatography showed that each was composed of three glycoproteins and that the major difference between the subcomponents was due to their constituent glycoproteins possessing different numbers of sialic acid residues. Combined analysis of the ESI-MS data and of the overall carbohydrate compositional data obtained by colorimetric procedures, leads to the composition of the glycan of each glycoprotein, and a combined methylation and 400 MHz H-NMR analysis of the alkaline cleaved glycans identified them as being of only the biantennary N-acetyllactosamine type. Taking into account the molecular mass, the carbohydrate content and structure it may be concluded that each of the constituent glycoproteins contain five N-glycosidically linked glycans.

Mass spectrometric composition, molecular mass and oxygen binding of Macrobdella decora hemoglobin and its tetramer and monomer subunits.

The hexagonal bilayer hemoglobin (Hb) of the leech Macrobdella decora has an equilibrium sedimentation mass of 3544(+/- 80) kDa. Maximum entropy analysis of the electrospray ionization mass spectra of the Hb show three groups of peaks: two peaks of equal intensity at approximately 17 kDa, A (16,770.1 Da) and B (16,841.9 Da); three peaks at approximately 24 kDa, C (24,340.1 Da), D (24,398.6 Da) and E (24,420.0 Da) with relative intensities of 1:6:3, respectively; and three peaks of equal intensities at approximately 33 kDa, F (32,586.0 Da), G (32,714.5 Da) and H (32,849.9 Da). Although reduction with dithiothreitol does not affect the masses of peaks A through E, the approximately 33 kDa peaks give rise to four new peaks at approximately 16 kDa, P (16,052.2 Da), Q (16,537.3 Da), R (16,666.7 Da) and S (16,792.9 Da), indicating that F, G and H represent disulfide-bonded dimers of globin chains, P + Q, P + R and P + S, respectively. The relative intensities of the three groups of peaks are (A + B) to (C + D + E) to (F + G + H) = 0.39:0.26:0.32, and the globin to linker ratio 0.71:0.29 is in good agreement with the ratio 0.72:0.28 obtained by HPLC. The largest functional subunit obtained by dissociation at pH 7 in 4 M urea, is a subunit lacking linker chains with apparent mass 63(+/- 3) kDa. The equilibrium sedimentation profile of this subunit is fitted best as a monomer-dimer-tetramer equilibrium, with association constants K1,2 = 365 l g-1 and K1,4 = 8.1 x 10(5) l3 g-3. A model of the Hb consisting of a hexagonal bilayer of 36 tetramer and 42 linker subunits provides a total mass and globin to linker ratio closest to the experimental values. Equilibrium O2 binding measurements of the native Hb and its tetramer and monomer subunits were carried out over the pH range 6.6 to 8.0 at 10 and 25 degrees C, and in the absence and presence of Na+, Mg2+ and Ca2+. The Hb exhibits a moderately high O2 affinity, P50 = 4.4 torr at pH 7.5 and 25 degrees C, a high cooperativity (n50 approximately 3) and a substantial Bohr effect, phi = delta log P50/delta pH = -0.38. The tetramer subunit has a higher affinity, lower cooperativity and smaller Bohr effect, 1.9 torr, 1.3 to 1.5 and -0.30, respectively. The monomer subunit has a much higher affinity (P50 = 0.29 torr) and no cooperativity or Bohr effect.(ABSTRACT TRUNCATED AT 400 WORDS)

Spectrum of beta-thalassaemia mutations in the population of Saudi Arabia.

A study of beta-thalassaemia alleles in 29 beta-thalassaemia homozygotes and HbS/beta-thalassaemia patients from western Saudi Arabia using the allele-specific priming technique of the polymerase chain reaction revealed a total of five mutations. The spectrum of mutations found in the western region is significantly different to that previously reported in the eastern region, and brings the total number of mutations observed in Saudi Arabian nationals to ten. A comparison of the types and frequencies of the Saudi Arabian mutations with those found in neighboring Arab countries is made.