Human TRP-1 Has Tyrosine Hydroxylase but no DOPA Oxidase Activity

Human TRP-1 has been immunopurified from normal human melanocytes cultured from black neonatal subjects and used to investigate the catalytic function of TRP-1 for the two substrates, L-tyrosine and L-DOPA. Immunopurified TRP-1 did not demonstrate DOPA staining on SDS/PAGE nor DOPA oxidase (DO) activity with either routine or modified assays. The purified TRP-1 also demonstrated no tyrosine hydroxylase (TH) activity using the routine Pomerantz assay. However, there was apparent TH activity exhibited by immunopurified TRP-1 under conditions with low tyrosine concentration (≤0.8 μCi/ml of ³H-tyrosine), prolonged incubation time (i.e., overnight) and in the absence of the cofactor L-DOPA. Using these latter specific conditions, TH activity was also detected in cell lysates from a tyrosinase-negative albino melanocyte line which exhibited no TH activity with the routine Pomerantz assay. In addition, TH activity under low substrate assay conditions was not exhibited in a melanocyte line derived from a TRP-1 deficient, Brown albino individual. However, the absence of TH in this Brown albino cell line could be compensated for by the addition of L-DOPA to the assay. These results suggested that TRP-1 has some tyrosine hydroxylase but no DOPA oxidase activity. We propose that one function of TRP-1 is to modulate tyrosinase activity by making DOPA available as a cofactor to perpetuate the initial steps in melanogenesis.     

Histological,Biochemical, and Ultrastructural Studies on Hyperpigmented Human Skin Xenografts

The mechanisms for hyperpigmentation observed in human cutaneous xenografts placed on athymic nude mice was investigated. Histologic, biochemical, histochemical, and ultrastructural examinations were performed on human skin prior to grafting and at various times ranging from 2 weeks to 30 weeks post-grafting (PG). Hyperpigmentation was macroscopically visible on the graft as early as 4–6 weeks. The number of Dopa-positive melanocytes per unit area was increased at 2 weeks PG and remained elevated until 20 weeks PG. The surface area of the melanocytes, a measure of the activity of the cells, also increased significantly and remained above the pre-grafting size throughout the study. Western blot analysis using tyrosinase specific antibody (αTy-SP) revealed the presence of tyrosinase exclusively in the grafted skin from 2 weeks to 12 weeks PG tested. Histological and ultrastructural observations revealed the presence of numerous dendritic melanocytes, indeterminant clear cells suggestive of Langerhans cells, and dermal melanophages. The results of this study suggest that the observed hyperpigmentation in grafted tissue is caused by an increase in the number of Dopa-positive melanocytes and probably from enhanced melanin production. Extracts of proteins from the xenografts exhibited prominent differences in low and high molecular proteins between pre- and post-grafted skin. Among them, the exclusive appearance of a protein doublet with apparent mw ～14 kDa was found in grafted skin, and subsequent studies indicate it has potent effects on melanocyte function.     

Population structure in Mediterranean islands and risk of genetic invasion in Culexpipiens L. (Diptera: Culicidae)

The mosquito Culex pipiens is subjected to organophosphate treatments in both Corsica and southern Sardinia, but the resistance gene A2-B2, which is currently in a worldwide expansion, has only reached Sardinia. In order to understand this situation, the genetic structure of populations sampled in Sardinia and Corsica was assessed using 15 isozymes. Two loci (HK1 and HK2) were not taken into account because of the possibility of selection. For trie other loci, statistical independence was not rejected for all possible pairs, and no deviation from Hardy-Weinberg expectations was apparent. Low but significant genie differentiation was present between Corsica and Sardinia, as well as between northern and southern Sardinia, despite a large number of effective migrants per generation. These results are discussed in the context of the high probability of extinction/recolonization of breeding sites, the flight migration ability of this mosquito, and the pleiotropic cost of insecticide resistances genes. It is concluded that A2-B2 resistance is unlikely to reach Corsica from southern Sardinia, unless accidental human transportation occurs.     

Cleavage Furrow Establishment--A Preliminary to Cylindrical Shape Change

The predictable pattern of cell shape changes characterizinganimal development must be a consequence of control mechanismsthat are at least analogous to those operating in dividing cells.When cells change shape by an internal mechanism, it is impliedthat they also contain systems that will activate and deactivatethe mechanism, localize it within the cell, insure proper timing,and impart the proper vectoral qualities. Experimental investigationsdesigned to elucidate similar processes in dividing cells revealthat the physical mechanism that accomplishes cytokinesis isestablished at or near the equatorial cell surface by the mitoticapparatus. The process is completed by late metaphase or earlyanaphase. In cleaving eggs a pair of asters can substitute forthe intact mitotic apparatus. The nature of the stimulus whichapparently passes from the mitotic apparatus to the surfaceis presently unknown. It moves toward the surface at about 6microns per minute and requires about 1 minute to establishthe mechanism. The resulting equatorial contractile activityis initially isotropic but becomes anisolropic at the beginningof visible constriction.     

Ultraviolet reflectance mediates pollinator visitation in Mimulus guttatus

Floral colors are widely believed to be an adaptation to attract pollinators. Recently, our understanding of floral reflectance has broadened to include colors that are beyond the spectrum that human eyes can perceive (such as ultraviolet (UV) reflectance), yet we still know relatively little about which plant species reflect UV light or its effectiveness in attracting pollinators. We investigated the effect of UV reflectance in Mimulus guttatus in a number of different populations in British Columbia, Canada. We found that M. guttatus had distinct regions of the corolla where UV light was reflected and absorbed. When we manipulated the degree of contrast between the reflection and absorption area, we found that pollinator visitation was severely disrupted, in terms of frequency and foraging patterns observed. Despite the bright yellow (bee‐green) coloration and visible nectar guides in M. guttatus, we conclude that UV reflectance is critical in pollinator attraction.     

Microsatellite markers developed from Theobroma cacao L. expressed sequence tags

Theobroma cacao L. expressed sequence tags (ESTs) were converted into useful genetic markers for fingerprinting individuals and genetic linkage mapping. Primers were designed to microsatellite‐containing ESTs. Twenty‐two T. cacao accessions, parents of various mapping populations segregating for disease resistance and crop yield characteristics, were tested. Twenty‐seven informative loci were discovered with 26 primer pairs. The number of detected alleles ranged from two to 11 and averaged 4.4 per locus. All 27 markers could be mapped into at least one of the existing F₁ or F₂ populations segregating for agronomically important traits.     

A new species of the marattialean fern Scolecopteris (Zenker) Millay from the uppermost Permian of Guizhou Province, south-western China

Several isolated marattialean synangia and sporangia are reported from coal balls collected from Coal Seam No.1 (C605) in the uppermost Permian Wangjiazhai Formation in Guizhou Province, south-western China. The synangia are radially symmetrical with diameters between 0.8 and 1.2 mm and are 1.7 mm long, consisting of 3–4 elongate sporangia that are fused basally, free distally and possess a pointed apex. The outer-facing sporangial wall is 4–5 cells thick and conspicuously differentiated. Spores are trilete, have a granular ornamentation and are nearly round equatorially with a diameter of 55–60 µm. Comparisons with other anatomically preserved Palaeozoic marattialean synangia from the Euramerican and Cathaysian floras permit their assignment to the genus of Scolecopteris (Zenker) Millay. In this species the thick, outer-facing sporangial walls and large trilete spores are features consistent with those of the Oliveri Group within Scolecopteris , a group that has previously been considered primitive within this genus. Distinctions from all other previously recognized species within the Oliveri Group lead to the creation of a new species, S. guizhouensis sp. nov. This species is the youngest of the reported species of Scolecopteris recognized from the Euramerican and Cathaysian floras, and provides important evidence on the organization of marattialean ferns from the Upper Permian strata of south China.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 151 , 279–288.     

Testing the Adaptive Significance of Acclimation: A Strong Inference Approach

Acclimation is a common phenotypic response to environmentalchange. Acclimation is often thought to enhance performanceand thus to be adaptive. This view has recently been formalizedas the "Beneficial Acclimation Hypothesis" and predicts thatindividuals acclimated to one environment perform better inthat environment than do individuals acclimated to a differentenvironment. Although Beneficial Acclimation is appealing andwidely supported, recent studies with E. coli and Drosophilahave challenged its general validity. Although these challengescould be dismissed as mere exceptions, they encourage a re-evaluationof the adaptive significance of acclimation. Our philosophicalapproach differs from that of most previous studies of acclimation,in which the prediction derived from a Beneficial Acclimationperspective (e.g., heat tolerance is positively correlated withacclimation temperature) is tested against the null hypothesis("single hypothesis approach"). Instead, we follow Huey andBerrigan (1996) in advocating a strong inference approach (sensuPlatt, 1964), which recognizes that Beneficial Acclimation isactually one of a set of competing hypotheses that make differentpredictions as to how developmental temperature influences thethermal sensitivity of adults ("developmental acclimation").Using this perspective, Huey and Berrigan proposed a factorialexperimental design (3 developmental by 3 adult temperatures)designed to discriminate among all competing hypotheses. Wenow derive a formal statistical model (ANOVA with orthogonalpolynomial contrasts) for this experimental design and use itto evaluate simultaneously the relative impact of each competinghypothesis. We then apply this model to several case studies(Drosophila, Volvox, Trichogramma), and we review also a recentstudy with E. coli. The influence of Beneficial Acclimationis supported (albeit often weakly) in most cases. Nevertheless,other hypotheses (especially the Optimal Developmental TemperatureHypothesis) often have a greater impact. Even so, however, BeneficialAcclimation usually predicts relative performance at extremetest temperatures. We conclude that, although rumors of itsdeath are premature, Beneficial Acclimation cannot be viewedas the dominant expectation, at least with regard to developmentaltemperature acclimation. Moreover, our findings reinforce theview that a strong inference approach provides a more comprehensiveportrait of complex biological responses than do single-hypothesisapproaches.