Male death resulting from hybridization between subspecies of the gypsy moth, Lymantria dispar.

We explored the origin of all-female broods resulting from male death in a Hokkaido population of Lymantria dispar through genetic crosses based on the earlier experiments done by Goldschmidt and by testing for the presence of endosymbionts that are known to cause male killing in some insect species. The mitochondrial DNA haplotypes of the all-female broods in Hokkaido were different from those of normal Hokkaido females and were the same as those widely distributed in Asia, including Tokyo (TK). Goldschmidt obtained all-female broods through backcrossing, that is, F1 females obtained by a cross between TK females (L. dispar japonica) and Hokkaido males (L. dispar praeterea) mated with Hokkaido males. He also obtained all-male broods by mating Hokkaido females with TK males. Goldschmidt inferred that female- and male-determining factors were weakest in the Hokkaido subspecies and stronger in the Honshu (TK) subspecies. According to his theory, the females of all-female broods mated with Honshu males should produce normal sex-ratio broods, whereas weaker Hokkaido sexes would be expected to disappear in F1 or F2 generations after crossing with the Honshu subspecies. We confirmed both of Goldschmidt's results: in the case of all-female broods mated with Honshu males, normal sex-ratio broods were produced, but we obtained only all-female broods in the Goldschmidt backcross and obtained an all-male brood in the F1 generation of a Hokkaido female crossed with a TK male. We found no endosymbionts in all-female broods by 4,'6-diamidino-2-phenylindole (DAPI) staining. Therefore, the all-female broods observed in L. dispar are caused by some incompatibilities between Honshu and Hokkaido subspecies.

A full-length cDNA resource for the pea aphid, Acyrthosiphon pisum.

Large collections of full-length cDNAs are important resources for genome annotation and functional genomics. We report the creation of a collection of 50 599 full-length cDNA clones from the pea aphid, Acyrthosiphon pisum. Sequencing from 5' and 3' ends of the clones generated 97 828 high-quality expressed sequence tags, representing approximately 9000 genes. These sequences were imported to AphidBase and are shown to play crucial roles in both automatic gene prediction and manual annotation. Our detailed analyses demonstrated that the full-length cDNAs can further improve gene models and can even identify novel genes that are not included in the current version of the official gene set. This full-length cDNA collection can be utilized for a wide variety of functional studies, serving as a community resource for the study of the functional genomics of the pea aphid.

Evasion, suppression and tolerance of Drosophila innate immunity by a male-killing Spiroplasma endosymbiont.

How endosymbiotic bacteria cope with host insect immunity is poorly understood. Here we report previously unknown aspects of immunity-mediated interactions between male-killing/non-male-killing spiroplasmas and Drosophila host. The male-killing spiroplasma tended to reduce constitutive expression levels of some antimicrobial peptide genes, while the non-male-killing spiroplasma did not. In mutant flies whose innate immunity is constitutively up-regulated, infection densities of the male-killing spiroplasma were significantly suppressed but managed to increase during the aging of adult flies, indicating that the male-killing spiroplasma is resistant to mounted immune attacks. These findings suggest that not only immune evasion but also immune suppression and tolerance are involved in the establishment and maintenance of the insect-microbe symbiotic association.

Early-stage dynamics of light-matter interaction leading to the insulator-to-metal transition in a charge ordered organic crystal.

Ultrafast dynamics of the light-matter interaction in a charge-ordered molecular insulator α-(BEDT-TTF)2I3 were studied by pump-probe spectroscopy using few-optical-cycle infrared pulses (pulse width 12 fs). Coherent oscillation of the correlated electrons and subsequent Fano destructive interference with intramolecular vibration were observed in time domain; the results indicated a crucial role for electron-electron interplay in the light-matter interaction leading to the photoinduced insulator-to-metal transition. The qualitative features of this correlated electron motion were reproduced by calculations based on exact many-electron-phonon wave functions.

Optical modulation of effective on-site coulomb energy for the Mott transition in an organic dimer insulator.

We report an optical modulation of the effective on-site Coulomb energy U on a dimer (U_{dimer}) for achieving the Mott insulator-to-metal transition in kappa-(BEDT-TTF)_{2}Cu[N(CN)_{2}]Br, as investigated by pump-probe spectroscopy. A reduction of U_{dimer} is optically induced by molecule displacement in the dimer under intradimer excitation. The mechanism of this metallization differs greatly from the photodoping-type mechanism reported previously. In contrast, a faster transition via the photodoping mechanism is detected for interdimer excitation. A metallic-domain-wall oscillation originating from the modulation of U_{dimer} was also observed near the critical end point of the Mott transition line.

Effects of host genotype against the expression of spiroplasma-induced male killing in Drosophila melanogaster.

Increasing attention has been paid to the maternally inherited microbes that are capable of manipulating the reproduction of their hosts for their own benefit. Although several studies have revealed that the host genotype can affect the intensity of the manipulation, the underlying genetic basis is poorly understood. Here, we examined the intensity of spiroplasma-induced male killing in various wild-type stocks of Drosophila melanogaster to clarify the genetic basis of the host factors responsible for the variation in the male-killing intensity. Among ten lines examined by mating experiments (that is, nuclear introgression), eight lines including Oregon-R and Canton-S were found to have nuclear factors that allowed strong expression of male killing. In contrast, the nuclear factors of the lines Sevelen and Hikone partially suppressed or remarkably retarded the expression of male killing. These results were confirmed by artificial transfer experiments of spiroplasma infection across the fly lines by means of microinjection. A series of mating experiments revealed that the nuclear factors acting against male killing were mainly located on autosomes in Sevelen and on the X chromosome in Hikone. In both lines, the suppressors were inferred to act maternally with a dominant effect. The nuclear factors of Sevelen and Hikone scarcely affected spiroplasma densities in reproductively active young insects, suggesting that the suppressors may act on the male-killing expression directly rather than through suppressing bacterial proliferation.

Role of host nutrition in symbiont regulation: impact of dietary nitrogen on proliferation of obligate and facultative bacterial endosymbionts of the pea aphid Acyrthosiphon pisum.

The impact of host nutrition on symbiont regulation in the pea aphid Acyrthosiphon pisum was investigated. The population density of the obligate symbiont Buchnera aphidicola positively correlated with dietary nitrogen levels. In contrast, the population density of the facultative symbiont Serratia symbiotica increased in aphids reared on low-nitrogen diets, indicating distinct regulatory mechanisms in the same insect host.

Rickettsia infection in natural leech populations.

Field-collected specimens of glossiphoniid leeches, Torix tagoi, Torix tukubana, Hemiclepsis marginata, and Hemiclepsis japonica, were surveyed for Rickettsia infection by using a diagnostic PCR assay. Rickettsia was detected in 96% (69/72) of T. tagoi, 83% (24/29) of T. tukubana, 29% (33/113) of H. marginata, and 0% (0/30) of H. japonica. The frequencies of Rickettsia infection were stably maintained in different seasons. In H. marginata and T. tukubana, distant local populations exhibited remarkably different frequencies of Rickettsia infection. Eggs carried by infected females of T. tagoi and H. marginata were all Rickettsia-positive, indicating nearly 100% vertical transmission. Analysis of 16S rDNA sequences revealed that phylogenetic relationship of the leech-associated Rickettsia reflected the specific and populational divisions of the host leeches. However, circumstantial lines of evidence strongly suggested that horizontal transmission of Rickettsia must have occurred in the ancestors of these leeches. In T. tagoi and T. tukubana, infected individuals were remarkably larger in size than uninfected individuals, wheras in H. marginata, infected and uninfected individuals were almost comparable in size. This study first provides information on ecological aspects of leech-bone endocellular bacteria of the genus Rickettsia. On the basis of these data, we discuss possible mechanisms whereby Rickettsia infection is maintained in natural populations of these leeches in the freshwater ecosystem.

Characterization of a facultative endosymbiotic bacterium of the pea aphid Acyrthosiphon pisum.

The pea aphid U-type symbiont (PAUS) was investigated to characterize its microbiological properties. Fluorescence in situ hybridization (FISH) and electron microscopy revealed that PAUS was a rod-shaped bacterium found in three different locations in the body of the pea aphid Acyrthosiphon pisum: sheath cells, secondary mycetocytes, and hemolymph. Artificial transfer experiments revealed that PAUS could establish stable infection and vertical transmission when introduced into uninfected pea aphids. When 28 aphid species collected in Japan were subjected to a diagnostic PCR assay, four species of the subfamily Aphidinae (Aphis citricola, Aphis nerii, Macrosiphum avenae, and Uroleucon giganteus) and a species of the subfamily Pemphiginae (Colopha kansugei) were identified to be PAUS-positive. The sporadic incidences of PAUS infection without reflecting the aphid phylogeny can be best explained by occasional horizontal transfers of the symbiont across aphid lineages.

Transmission of symbiotic bacteria Buchnera to parthenogenetic embryos in the aphid Acyrthosiphon pisum (Hemiptera: Aphidoidea).

All phloem-feeding aphids have an absolute requirement for their primary bacterial symbionts Buchnera sp. The bacteria are transmitted vertically to either embryos in the viviparous morph or to eggs in the oviparous morph, with the implication that the symbiont population regularly passes through a genetic 'bottleneck', i.e. only a small proportion of the maternal symbiont population is transmitted to offspring. In this paper, we visualise this process in viviparous aphids using a specific immunolabelling technique for Buchnera. The images show a stream of bacteria originating from a single mycetocyte and entering the embryo, possibly via a membranous conduit, and individual bacterial cells free in the haemocoel of the aphid. Staining within the embryo blastoderm suggests over expression of antigen, perhaps indicative of rapid bacterial division immediately following infection.

Maintenance of soldier-producing aphids on an artificial diet.

In our efforts to understand the biology of soldier-producing aphids, we attempted to maintain them in the laboratory using a chemically defined artificial diet. The ability of 16 species from the subfamilies Eriosomatinae and Hormaphidinae, most of which are soldier-producing species, to survive on the artificial diet was examined. Some species neither fed nor grew on the diet, whereas other species accepted the diet, grew to some extent, and managed to produce a small number of short-lived offspring. Although they performed poorly on the diet in general, aphid performance was correlated with the stage in the life cycle and the developmental stadium in that aphids of the gall generation tended to accept the diet and survive on it, whereas aphids of the non-gall generation did not. Also, old insects tended to perform better on the diet than young nymphs. Notably, only one species, Tuberaphis styraci, a gall-forming aphid that produces 2nd instar sterile soldier, showed good performance on the diet. Insects collected from galls (generation G0) survived on the diet, grew well, and produced many progeny. Three successive generations (G1, G2 and G3) were produced on the diet. Developmental period, adult body size, and age of first reproduction were almost constant through G0, G1 and G2 whereas fecundity, adult longevity and daily offspring production declined as the generations proceeded. These results are comparable to previous studies in which pest aphids have been maintained on similar artificial diets for several generations. Therefore, it is suggested that the artificial-diet rearing system will provide a useful tool to investigate various biological aspects of the soldier-producing eusocial aphid, T. styraci.

Secondary intracellular symbiotic bacteria in aphids of the genus Yamatocallis (Homoptera: Aphididae: Drepanosiphinae).

A novel secondary intracellular symbiotic bacterium from aphids of the genus Yamatocallis (subfamily Drepanosiphinae) was characterized by using molecular phylogenetic analysis, in situ hybridization, and diagnostic PCR detection. In the aphid tissues, this bacterium (tentatively designated YSMS [Yamatocallis secondary mycetocyte symbiont]) was found specifically in large cells surrounded by primary mycetocytes harboring Buchnera cells. Of nine drepanosiphine aphids examined, YSMS was detected in only two species of the same genus, Yamatocallis tokyoensis and Yamatocallis hirayamae. In natural populations of these aphids, YSMS was present in 100% of the individuals. Phylogenetic analysis based on 16S ribosomal DNA (rDNA) sequences demonstrated that YSMS of Y. tokyoensis and Y. hirayamae constitute a distinct and isolated clade in the gamma subdivision of the class Proteobacteria. No 16S rDNA sequences of secondary endosymbionts characterized so far from other aphids showed phylogenetic affinity to YSMS. Based on these results, I suggest that YSMS was acquired by an ancestor of the genus Yamatocallis and has been conserved throughout the evolution of the lineage. By using the nucleotide substitution rate for 16S rDNA of Buchnera spp., the time of acquisition of YSMS was estimated to be about 13 to 26 million years ago, in the Miocene epoch of the Tertiary period.

Evolutionary dynamics of multiple group I introns in nuclear ribosomal RNA genes of endoparasitic fungi of the genus Cordyceps.

A large number of group I introns were discovered in coding regions of small and large subunits of nuclear ribosomal RNA genes (SSU rDNA and LSU rDNA) in ascomycetous fungi of the genus CORDYCEPS: From 28 representatives of the genus, we identified in total 69 group I introns which were inserted at any of four specific sites in SSU rDNA and four specific sites in LSU rDNA. These group I introns reached sizes of up to 510 bp, occurred in up to eight sites in the same organism, and belonged to either subgroup IB3 or subgroup IC1 based on their sequence and structure. Introns inserted at the same site were closely related to each other among Cordyceps fungi, whereas introns inserted at different sites were phylogenetically distinct even in the same species. Mapped on the host phylogeny, the group I introns were generally not restricted to a particular lineage, but, rather, widely and sporadically distributed among distinct lineages. When the phylogenetic relationships of introns inserted at the same site were compared with the phylogeny of their hosts, the topologies were generally significantly congruent to each other. From these results, the evolutionary dynamics of multiple group I introns in Cordyceps fungi was inferred as follows: (1) most of the group I introns were already present at the eight sites in SSU and LSU rDNAs of the ancestor of the genus Cordyceps; (2) the introns have principally been immobile and vertically transmitted throughout speciation and diversification of Cordyceps fungi, which resulted in the phylogenetic congruence between the introns at the same site and their hosts; (3) in the course of vertical transmission, the introns have repeatedly been lost in a number of lineages independently, which has led to the present sporadic phylogenetic distribution of the introns; and (4) a few acquisitions of new introns, presumably through horizontal transmission, were identified in the evolutionary history of the genus Cordyceps, while no transpositions were detected. Losses of group I introns in SSU rDNA have occurred at least 27 times in the evolutionary course of the 28 Cordyceps members.

Genetically distinct populations in an Asian soldier-producing aphid, Pseudoregma bambucicola (Homoptera: Aphididae), identified by DNA fingerprinting and molecular phylogenetic analysis.

To estimate genetic structure of a soldier-producing aphid, Pseudoregma bambucicola, samples from natural populations throughout southeastern Asia were analyzed by a DNA fingerprinting technique. We unexpectedly found that P. bambucicola comprises two geographic groups, the northern group and the southern group, which are genetically distinct from each other but morphologically almost indistinguishable. Molecular phylogenetic and statistical analyses based on mitochondrial ribosomal DNA sequences demonstrated that the northern and southern groups of P. bambucicola are not closely related but constitute distinct lineages in the genus Pseudoregma. Detailed morphological reexamination revealed that the two groups could be distinguished by the number of setae on the 8th abdominal tergite of 1st instar nymphs and soldiers. From these results, it was suggested that P. bambucicola should be divided into two species. The northern group from Japan, Taiwan, Hong Kong, and northern Vietnam retains the name P. bambucicola, whereas we suggest that the name P. carolinensis (R. Takahashi, 1941, Tenthredo 3, 208-220) should be used for the southern group from Thailand, Malay Peninsula, Java, Irian Jaya, and Micronesia. The morphological resemblance between P. bambucicola and P. carolinensis might be due to shared ancestral characters of the genus Pseudoregma.

Spiroplasma symbiont of the pea aphid, Acyrthosiphon pisum (Insecta: Homoptera).

From a laboratory strain of the pea aphid, Acyrthosiphon pisum, we discovered a previously unknown facultative endosymbiotic bacterium. Molecular phylogenetic analysis based on 16S ribosomal DNA revealed that the bacterium is a member of the genus Spiroplasma. The Spiroplasma organism showed stable vertical transmission through successive generations of the host. Injection of hemolymph from infected insects into uninfected insects established a stable infection in the recipients. The Spiroplasma symbiont exhibited negative effects on growth, reproduction, and longevity of the host, particularly in older adults. Of 58 clonal strains of A. pisum established from natural populations in central Japan, 4 strains possessed the Spiroplasma organism.

[Rupture of renal pelvis due to urolithiasis: report of two cases].

Two cases of renal pelvis rupture caused by urolithiasis are reported. The first case was in a 62-year-old male who had left flank pain. Urological examination including drip infusion pyelography (DIP) and abdominal computed tomography (CT) revealed a spontaneous rupture of the left renal pelvis. The rupture was assumed to have been caused by a small ureteral stone. The stone passed spontaneously, and the extravasation disappeared with conservative therapy. The second case was in a 84-year-old male who was admitted with high fever. Urological examination including DIP and abdominal CT showed a suspected rupture of the left renal pelvis due to a renal stone at the ureteropelvic junction. Percutaneous nephrostomy was performed and antegrade pyelography showed extravasation from the left renal pelvis. Extravasation disappeared on the 12th postoperative day. Extracorporeal shock wave lithotripsy was performed three times. The renal stone was completely discharged and the nephrostomy tube was removed successfully.

[No relation between angiotensin-converting enzyme (ACE) inhibitor-induced cough and ACE gene polymorphism, plasma bradykinin, substance P and ACE inhibitor concentration in Japanese patients].

Persistent dry cough is well known as the most common side-effect of angiotensin-converting enzyme (ACE) inhibitors. We examined the relationship between a cough and ACE gene polymorphism, plasma bradykinin (BK), substance P (SP) and ACE inhibitor concentrations in patients with hypertension or chronic nephritis. First, ACE genotyping was carried out in 96 patients, 42 with coughs and 54 without coughs, which had been treated with various kinds of ACE inhibitors. However, no significant difference in the ACE genotypes was observed between the two groups. Second, the plasma concentrations of BK, SP and ACE inhibitor were measured in 12 patients, which were treated with trandolapril at a daily dose of 1 mg for 4-33 weeks. In 3 patients, the cough was induced during the trandolapril therapy, while it was induced not in 9 patients. The plasma levels of BK and SP did not significantly change after trandolapril administration in the patients with and without coughs. Between the two groups, there were no significant differences in the plasma levels of BK and SP either before or after the trandolapril therapy. Also the plasma concentrations of trandolapril and trandolaprilat, the active metabolite of trandolapril, did not significantly differ between the two groups. These results suggest that there is no significant relationship between the ACE inhibitor-induced cough and ACE gene polymorphism, plasma BK, SP and ACE inhibitor concentrations in patients with hypertension or chronic nephritis.

[Alkaline phosphatase].

High levels of alkaline phosphatase activity are characteristic of bone disease with increased osteoblastic activity, hepatobiliary disease with partial or complete biliary obstruction and transient hyperphosphatasemia in children. A gamma-glutamyl transpeptidase is often useful to differentiate whether an elevated alkaline phosphatase originates in the bones or the liver. A normal gamma-glutamyl transpeptidase suggests bone origin. A serum alkaline phosphatase isozyme pattern by electrophoresis is also necessary to differentiate various pathological states.

[Two cases of abdominal masses caused by foreign bodies which were preoperatively diagnosed as urachal abscess].

A 61-year-old man and a 59-year-old woman were referred to our hospital because of lower abdominal pain and discomfort, pollakisuria and a lower abdominal mass. In both patients, radiological studies and cystoscopy caused us to suspect a urachal abscess. We performed operations transperitoneally. In the male patient, fish bones were detected between the mass and ileum. A partial cystectomy was performed on the female patient, and the histological diagnosis was Sparganosis mansoni. In both cases, it was very difficult to make a correct diagnosis before the operations, but surgical treatment was successfully performed.