Geographic body size variation in the periodical cicadas Magicicada: implications for life cycle divergence and local adaptation.

Seven species in three species groups (Decim, Cassini and Decula) of periodical cicadas (Magicicada) occupy a wide latitudinal range in the eastern United States. To clarify how adult body size, a key trait affecting fitness, varies geographically with climate conditions and life cycle, we analysed the relationships of population mean head width to geographic variables (latitude, longitude, altitude), habitat annual mean temperature (AMT), life cycle and species differences. Within species, body size was larger in females than males and decreased with increasing latitude (and decreasing habitat AMT), following the converse Bergmann's rule. For the pair of recently diverged 13- and 17-year species in each group, 13-year cicadas were equal in size or slightly smaller on average than their 17-year counterparts despite their shorter developmental time. This fact suggests that, under the same climatic conditions, 17-year cicadas have lowered growth rates compared to their 13-years counterparts, allowing 13-year cicadas with faster growth rates to achieve body sizes equivalent to those of their 17-year counterparts at the same locations. However, in the Decim group, which includes two 13-year species, the more southerly, anciently diverged 13-year species (Magicicada tredecim) was characterized by a larger body size than the other, more northerly 13- and 17-year species, suggesting that local adaptation in warmer habitats may ultimately lead to evolution of larger body sizes. Our results demonstrate how geographic clines in body size may be maintained in sister species possessing different life cycles.

Exaggerated male genitalia intensify interspecific reproductive interference by damaging heterospecific female genitalia.

Male-male competition over fertilization can select for harmful male genital structures that reduce the fitness of their mates, if the structures increase the male's fertilization success. During secondary contact between two allopatrically formed, closely related species, harmful male genitalia may also reduce the fitness of heterospecific females given interspecific copulation. We performed a laboratory experiment to determine whether the extent of genital spine exaggeration in Callosobruchus chinensis males affects the fitness of C. maculatus females by injuring their reproductive organs. We found that males with more exaggerated genital spines were more likely to injure the females via interspecific copulation and that the genital injury translated into fecundity loss. Thus, as predicted, reproductive interference by C. chinensis males on C. maculatus females is mediated by exaggeration of the genital spine, which is the evolutionary consequence of intraspecific male-male competition. Harmful male traits, such as genital spines, might generally affect the extent of interaction between closely related species.

Modification of a melanoma discrimination index derived from hyperspectral data: a clinical trial conducted in 2 centers between March 2011 and December 2013.

BACKGROUND: The morphology of pigmented skin lesions (PSLs) is predominantly a result of varying concentrations and distributions of pigmented molecules such as melanin and hemoglobin. Based on these differences and the fact that their information is contained in cutaneous spectra, a hyperspectral imager (HSI) for pigmented melanoma and a single discrimination index derived from the resultant hyperspectral data are proposed. OBJECTIVE: To develop and evaluate a new discrimination index for melanomas, compared to the previous index. METHODS: A HSI, which is convenient for both patients and clinicians, was newly developed and used in a clinical trial conducted in 2 centers with 80 patients with primary lesions and 17 volunteers between March 2011 and December 2013. There were 24 melanomas and 110 other PSLs. A previously proposed discrimination index was used without modifications. A new index, which emphasized the essential features of melanoma, was proposed, and its performance was examined. For each index, a threshold value was set to minimize the average value of the false positive and false negative fractions. The performances of both indices were compared. RESULTS: The sensitivity and specificity of the old index were 75% and 97%, respectively, while those of the new index were 96% and 87%. CONCLUSION: The new index had a higher sensitivity and adequate specificity, indicating that it is more useful than the old index.

How the length of genital parts affects copulation performance in a carabid beetle: implications for correlated genital evolution between the sexes.

To identify factors leading to the correlated evolution of exaggerated male and female genitalia, we studied the effects of the variable dimensions of corresponding functional genital parts (male copulatory piece and female vaginal appendix) on copulatory performance in the polygamous carabid beetle Carabus (Ohomopterus) maiyasanus. We used mating pairs of individuals from two populations to increase the variances in genital dimensions and determined the copulation performance (insemination and spermatophore replacement, and copulation time) in single- and double-mating situations. In single mating, insemination success was not affected by genital dimensions, although the copulation time was significantly shorter when the male aedeagus was longer. In the double-mating experiment, insemination and replacement of spermatophores by the second male succeeded more frequently when the copulatory piece was shorter and the vaginal appendix was longer, and when the difference between the length of the copulatory piece and the vaginal appendix was smaller. Thus, a matching of the corresponding genital parts between the sexes increases the male's reproductive success in sperm competition, but elongation of the copulatory piece cannot be explained simply by the improvement in male reproductive success. We discuss possible factors for the elongation of genital parts in terms of sexual conflict and reproductive interference through interspecific copulation.

Hyperspectroscopic imager for baby fibers.

Hyperspectral imaging system for diagnosing digestive diseases was newly developed in order to obtain information on pathology beyond morphology of lesions. In order to guide light reflected from a lesion, a baby fiber, which can be inserted in a forceps channel of the electronic endoscope, was also developed. The performance of the system was evaluated by animal experiment. Obtained hyperspectral data were found to have sufficient quality endurable to practical use. Harmful phenomena to a living body were not observed within the experiment. It was considered from the animal experiment that the present system could be practically used for humans.

Effects of lower body positive pressure on cardiovascular responses during walking in elderly women.

This study was undertaken to investigate the effects of lower body positive pressure (LBPP) on cardiovascular responses during a 15-min walking trial in young (22.1+/-0.4 years) and elderly women (67.8+/-1.1 years). The application of 20 mm Hg LBPP reduced ground reaction forces by 31.2+/-0.5 kgw in both groups. We hypothesized that cardiovascular responses to LBPP during walking were different between the young and elderly subjects. Applying 20 mm Hg of LBPP increased diastolic and mean blood pressure but not systolic blood pressure in both groups. LBPP-induced reduction in heart rate (HR) occurred more quickly in the young group compared to the elderly group (p<0.05). Applying LBPP also decreased double product (systolic blood pressure x HR) in both groups, suggesting that LBPP reduces myocardial oxygen consumption during exercise. These results suggest that heart rate responses to LBPP during exercise vary with increasing age.

Quantitative genetic analysis of subspecific differences in body shape in the snail-feeding carabid beetle Damaster blaptoides.

A dimorphic pattern of macrocephalic (wide, short) and stenocephalic (narrow, long) body shapes is observed in snail-feeding carabid beetles globally. The former exhibits high performance in crushing snail shells with powerful jaws, whereas the latter specializes in eating snails' soft body directly by inserting the head into the shell. In the snail-feeding species Damaster blaptoides, the subspecies D. b. capito has a wide, short forebody, and D. b. fortunei has a narrow, long forebody. They exhibit distinct morphologies despite their geographic and phylogenetic proximity. To examine the genetic basis of the morphological differences between these two subspecies, we conducted quantitative genetic analyses by crossing these subspecies and producing F(1) and backcross hybrids. The hybrids had body shapes intermediate between the parental subspecies. The variation between wide, short and narrow, long forebodies was based on negative genetic correlations between width and length of the head and thorax. Between one and eight genetic factors were involved in the morphological differences between subspecies. We suggest that the morphological integration of forebody parts in a small number of loci has facilitated the marked morphological diversification between subspecies of D. blaptoides.

Genetic basis of male colour dimorphism in a Lake Tanganyika cichlid fish.

Phenotypic polymorphisms can be applied to study the micro-evolutionary forces that maintain genetic variation and can mediate speciation, but it can be difficult to determine the genetic basis of polymorphisms. Recently, restriction-site-associated DNA (RAD) sequencing has become popular, which can easily produce multiple single nucleotide polymorphisms from whole genomes. Here, we combined RAD sequencing, allele-specific PCR and Sanger sequencing to determine the genetic basis underlying male colour dimorphism of a Lake Tanganyika cichlid fish, Cyprichromis leptosoma. Our analyses using both a cross-family (two parents and 12 F2 males) and 64 wild individuals do not contradict a hypothesis that two alleles of one-locus control male colour dimorphism. Also, the locus may be located on a genome region that experiences reduced levels of recombination. Although more analyses will be needed to conclude these findings, this study is the first to suggest the genetic basis of a colour polymorphism using RAD sequencing.

Genetic differences and phenotypic plasticity in body size between high- and low-altitude populations of the ground beetle Carabus tosanus.

The body size of a univoltine carabid beetle Carabus tosanus on Shikoku Island, Japan, was clearly smaller in higher-altitude populations (subspecies), which possibly represents incipient speciation. To explore the determinants of altitudinal differences in body size in this species, we studied the degree of phenotypic plasticity by conducting rearing experiments at two constant temperatures and examined genetic differences through interpopulation crosses. At 15 °C, C. tosanus had a longer developmental period and a shorter adult body than at 20 °C. Nevertheless, variation in body size due to temperature effects (phenotypic plasticity) was small compared to the interpopulation differences, which suggests substantial genetic differences between populations (subspecies) at different altitudes. In F(1) offspring from crosses between a low-altitude (subspecies tosanus) and a high-altitude population (subspecies ishizuchianus), adult body length was affected by the genotypes of both parents, with an interaction effect of parental genotype and offspring sex. Further analyses revealed that adult body length was affected by sex-linked factors in addition to autosomal factors. These genetic differences in body size may have resulted from adaptations to different altitudes and may be important for the process of incipient speciation because body size differences could contribute to premating reproductive isolation.

Divergence of ovipositor length and egg shape in a brood parasitic bitterling fish through the use of different mussel hosts.

Bitterling fishes deposit their eggs on the gills of living mussels using a long ovipositor. We examined whether ovipositor length (OL) and egg shape correlated with differences in host mussel species in the family Unionidae among populations of the tabira bitterling (Acheilognathus tabira) in Japan. Bitterling populations that use mussels in the sub-family Anodontinae possessed longer ovipositors and more elongated eggs than those using mussels of Unioninae, as expected from the difference in host size between the sub-families (anodontine mussels are larger than unionine mussels). Based on a robust phylogeny of A. tabira populations, we demonstrated that the evolution of both OL and egg shape were correlated with host differences, but not with each other, suggesting that these traits have been selected for independently. Our study demonstrates how adaptive traits for brood parasitism may diverge with host shift due to different host availability and/or interspecific competition for hosts.

Melanoma screening system using hyperspectral imager attached to imaging fiberscope.

Early detection and proper excision of the primary lesions of melanoma are crucial for reducing melanoma-related deaths. In order to support the early detection of melanoma, melanoma screening systems have been extensively studied and developed. Recently we have proposed a melanoma discrimination index derived from hyperspectral data (HSD) in the visible-near infrared wavelength region. The index represents variegation in spectra over a lesion and works well in discriminating melanoma from other pigmented lesions. However the previous hyperspectral imager did not have an enough allowance for measurement of lesions. To overcome the problem with it, we have developed a hyperspectral imager attached to imaging fiberscope. This equipment has been able to accumulate HSD in a view field of φ40 mm within about 10 seconds, from which the above-mentioned melanoma discrimination index has been calculated. Performance of the system has been studied in nine cases of melanoma and 18 cases of non-melanoma, obtained from patients and volunteers, all of whom were Japanese. The index has achieved a sensitivity of 100 % and a specificity of 94.4 %.

Rapid diversification of male genitalia and mating strategies in Ohomopterus ground beetles.

We analysed evolutionary diversification and covariation in male genitalia and four mating traits related to sexual selection, i.e. testis size, spermatophore size, copulation duration and post-copulatory guarding duration, in Ohomopterus ground beetles using phylogenetically independent contrasts. Male genital size and mating duration have evolved more rapidly than body size and the other traits studied. Male genital size was negatively correlated with copulation duration, suggesting that elongated male genitalia may enable decreased time investment in a single copulation because it is more effective at facilitating spermatophore deposition. Male genital size was positively correlated with spermatophore size, suggesting coevolution between offensive and defensive male mating tactics because the elongated male genitalia may be advantageous in displacement of rivals' plug-like spermatophores, and decreased mating duration may intensify sperm competition. Thus, the remarkable diversity of male genitalia in Ohomopterus may have been facilitated by the interplay between inter- and intrasexual selection processes.

The genetic basis of interspecific differences in genital morphology of closely related carabid beetles.

Marked diversification of genital morphology is common in internally fertilizing animals. Although sexual selection may be the primary process controlling genital evolution, factors promoting genital evolution are controversial, and the genetic background of genital morphology is poorly understood. We analyzed the genetic basis of species-specific genital morphologies in carabid beetles of the subgenus Ohomopterus (genus Carabus, Carabidae) using two parapatric species with hybrid zones. Biometric analyses on experimental F(1) and backcross populations revealed that inheritance of genital morphology is polygenic. Applying Lande's modification of the Castle-Wright estimator to population means and variances to estimate the minimum number of genes involved, we found that a relatively small number of loci is responsible for species differences in genital morphology. In addition, joint-scaling tests indicated that the additive genetic effect accounts for most interspecific differences in genital traits, but dominance and epistatic genetic effects also play roles. Overall, the genetic basis of male and female genitalia is fairly simple, enabling these traits to respond quickly to selection pressures and to diverge rapidly. Our results provide insight into the diversification of genital morphology in carabid beetles, and will hopefully stimulate further studies on the genetic basis of genitalia, such as mapping of quantitative trait loci affecting species-specific genital morphology.

Species status and phylogeography of two closely related Coptolabrus species (Coleoptera: Carabidae) in South Korea inferred from mitochondrial and nuclear gene sequences.

We investigated the species status and intraspecific phylogeography in South Korea of two ground beetle species, Coptolabrus jankowskii and Coptolabrus smaragdinus (Coleoptera: Carabidae), using statistical parsimony networks and nested clade analyses based on sequences from the mitochondrial cytochrome oxidase subunit I (COI) and nuclear phosphoenolpyruvate carboxykinase (PepCK) and wingless (Wg) genes. Although traditional parsimony tree construction generally failed to resolve interspecific relationships and construct biologically meaningful genealogies, analysis using statistical parsimony networks yielded statistically significant inter- and intraspecific genealogical structures. We found that although these two species represent a notable case of trans-species polymorphisms in both mitochondrial and nuclear gene sequences, their status as separate species was evidenced by the nonrandom association between species and nested clades at various nesting levels. The exceptional occurrence of shared identical or very similar COI sequences was considered to be the result of introgressive hybridization. In addition, range expansion and fragmentation events across the Korean Peninsula and adjacent islands were inferred from nested clade phylogeographical analyses. The COI gene revealed the geographical divergence of major eastern and western clades and historical biogeographical events within each major clade, whereas the nuclear PepCK gene, which did not reveal corresponding east-west clades, indicated past fragmentation and range expansion across wide areas that may have been the result of older biogeographical events. Thus, phylogeographical inferences drawn from analyses of mitochondrial and nuclear genes can reveal different and potentially complementary information about phylogeographical processes.

Evolutionary history of coastal tiger beetles in Japan based on a comparative phylogeography of four species.

To reveal the phylogeographical patterns of four species of coastal tiger beetles in Japan (Lophyridia angulata, Abroscelis anchoralis, Cicindela lewisii and Chaetodera laetescripta), we conducted phylogenetic and nested clade analysis (NCA) using the mitochondrial DNA sequences of two loci (COI and 16S rRNA), with specimens sampled from Japan and neighbouring countries. Abroscelis anchoralis and L. angulata have similar disjunct distributions in Japan. The NCA indicated past fragmentation involving three isolated areas of A. anchoralis. In contrast, local populations of L. angulata in Japan shared the same haplotype, indicating recent vicariance. Co-occurrence of haplotypes from several divergent clades in Japanese populations of Ch. laetescripta suggested ancient vicariance and subsequent intermixing of local populations. The tree topology of C. lewisii, with shallow branches and little geographical segregation of haplotypes between Japan and Korea or within Japan, suggested that the Japanese population was segregated from the Korean population only recently. Restricted gene flow, with isolation by distance, was inferred for various geographical associations of haplotypes for coastal tiger beetles in the NCA. Based on these phylogeographical patterns, coupled with a molecular clock approach, the evolutionary history of four species of coastal tiger beetles was deduced, with the additional consideration of the competitive relationships among those species. We also discuss the conservation of highly localized A. anchoralis populations in Japan, using the concept of evolutionarily significant units.

Extensive trans-species mitochondrial polymorphisms in the carabid beetles Carabus subgenus Ohomopterus caused by repeated introgressive hybridization.

To study the potential importance of introgressive hybridization to the evolutionary diversification of a carabid beetle lineage, we studied intraspecific and trans-species polymorphisms in the mitochondrial NADH dehydrogenase subunit 5 (ND5) gene sequence (1083 bp) in four species of the subgenus Ohomopterus (genus Carabus) in central and eastern Honshu, Japan. Of the four species, C. insulicola is parapatric with the other three, and can hybridize naturally with at least two. This species possesses two haplotypes of remote lineages. We classified ND5 haplotypes using polymerase chain reaction-restriction fragment length polymorphism with TaqI endonuclease for 524 specimens, and sequenced 143 samples. Analysis revealed that each species was polyphyletic in its mitochondrial DNA phylogeny, representing a marked case of trans-species polymorphism. Recent one-way introgression of mitochondria from C. arrowianus nakamurai to C. insulicola, and from C. insulicola to C. esakii, was inferred from the frequency of identical sequences between these species and from direct evidence of hybridization in their contact zones. Other intraspecific polymorphisms in the four species may be due to undetected introgressive hybridization (e.g. C. insulicola to C. maiyasanus) or from stochastic lineage sorting of ancestral polymorphisms. This beetle group has a genital lock-and-key system, with species-specific or subspecies-specific genital morphology that may act as a barrier to hybridization. However, our results demonstrate that introgressive hybridization has occurred multiple times, at least for mitochondria, despite differences among, and stability within, morphological characters that distinguish local populations. Thus, hybridization and introgression could have been key processes in the evolutionary diversification of Ohomopterus.

Incongruence of mitochondrial and nuclear gene trees in the Carabid beetles Ohomopterus.

We studied the molecular phylogeny of the carabid subgenus Ohomopterus (genus Carabus), using two mitochondrial (mt) DNA regions (16SrRNA and NADH dehydrogenase subunit 5) and three nuclear DNA regions (wingless, phosphoenolpyruvate carboxykinase, and an anonymous locus). We revisited the previously reported incongruence between the distribution of mtDNA markers and morphologically defined species (Su et al., 1996; J. Mol. Evol. 43:662-671), which those authors attributed to "type switching", a concerted change in many morphological characters that results in the repeated evolution of a particular morphological type. Our mtDNA gene tree obtained from 44 individuals representing all 15 currently recognized species of Ohomopterus revealed that haplotypes isolated from individuals of a single "species" were frequently separated into distant clades, confirming the previous report. The three nuclear markers generally conformed better-with the morphologically defined species than did the mitochondrial markers. The phylogenetic signal in mtDNA and nuclear DNA data differed strongly, and these two partitions were significantly incongruent with each other according to the incongruence length difference test of Farris et al. (1994; Cladistics 10:315-320), although the three nuclear partitions were not homogeneous either. Our results did not support the type-switching hypothesis that had been proposed to fit the morphological data to the mitochondrial gene tree: The incongruence of the mtDNA tree with other nuclear markers indicates that the mtDNA-based tree does not reflect species history any better than the morphological data do. Incongruence of gene trees in Ohomopterus may have been promoted by the complex processes of geographic isolation and hybridization in the Japanese Archipelago that have led to occasional gene flow and recombination between separated entities. The occurrence of reticulate patterns in this group is intriguing, because species of Ohomopterus exhibit extremely divergent genitalic structures that represent a highly efficient reproductive isolation mechanism.

Personality assessment of homeless adults as a tool for service planning.

The psychiatric status of homeless adults has been described primarily in terms of Axis I disorders. By adding a subset of the Personality Assessment Inventory, this study tests the feasibility and usefulness of a brief, self-administered questionnaire to obtain scores on several dimensions of personality. Cluster analysis sorted 112 tested subjects into four groups characterized by distinct profiles. Two of these were characterized by extreme scores on pathological dimensions of personality (borderline features, antisocial traits, and aggressivity) and differed primarily on the dimension of suicidality. The third reflected moderate levels of personality dysfunction and the fourth did not deviate from adult nonclinical norms. The validity of the clusters was supported by demographic, background, and diagnostic subgroup differences. Brief personality assessment can be a cost-effective approach to matching services with clinical needs of homeless adults by attending to interpersonal dimensions that will likely affect service provision.

Physical and biological attributes of water channels utilized by Culex pipiens pallens immatures in Saga City, southwest Japan.

Physical and biological attributes of water channels utilized by immatures of Culex pipiens pallens were studied in Saga City, Japan. Water in mosquito-productive segments generally was stagnant or slowly running (mean < 5 cm/sec), with low and fluctuating dissolved oxygen concentrations and high electric conductivity. Water flow > 20 cm/sec was considered necessary for prevention of mosquito breeding. The aquatic fauna in mosquito-productive segments was composed of taxa tolerable to polluted water. Adult Odonata were more diverse in segments with emergent vegetation irrespective of physical attributes of channel water. Fish diversity was higher in mosquito-free segments. Twenty-three fish species were confirmed in the creek networks. Temporary flooding did not flush mosquito immatures from mosquito-productive segments, indicating the high stability of those segments as mosquito immature habitats.