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.

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.

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.

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.

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.

Isolation and characterization of proteoglycans from human nonepithelial tumors.

Proteoglycans (PGs) and glycosaminoglycans (GAGs) were identified in myogenic and fibrogenic tumors. More PGs containing mainly chondroitin sulfate could be detected in malignant tumors (leiomyosarcomas) than in benign tumors (leiomyomas and fibromas). Two groups of PGs were detected in the malignant tumors by ion-exchange chromatography and gel chromatography. One group was a large molecule with chondroitin sulfate side chains, seemingly composed of two or more subpopulations that were eluted from Sepharose CL-4B with a kav of 0.45. After removal of GAG side chains from the PG by chondroitinase AC digestion, core molecules with molecular weights greater than 200,000 were obtained. Another PG detected was a fraction of small PG eluted from Sepharose CL-4B with a kav of 0.45. It consisted of a core molecule with a molecular weight approximately equal to 48,000 and GAG side chains containing chondroitin sulfate-dermatan sulfate hybrids. The mixed sequence of L-iduronic acid with D-glucuronic acid in the same GAG chain was demonstrated by the formation of a small proportion of tetrasaccharide after chondroitinase AC digestion. In the benign tumors, the large PG was found only in very small amounts, and PG detected was composed mainly of the small one eluted from Sepharose CL-4B with a kav of 0.45. Its core protein had a molecular weight of approximately equal to 46,000, which was similar to that of small PG obtained from leiomyosarcomas, but its GAG side chains were composed mainly of dermatan sulfate containing small amounts of glucuronic acid. The results suggest that the core molecules of small PGs from both benign and malignant tumors are the products of the same gene but that they are processed in a different manner to form proteoglycans with different types of GAG chains.

Effects of beta-D-xylosides on proliferation and matrix formation of adherent fibroblastic cells in mouse bone marrow culture.

Mouse bone marrow cells were seeded on small pieces of cover glasses placed in culture dishes, and after 3 days, the pieces of glass to which only small spindle cells adhered were transferred to another dish containing fresh medium. The adherent small spindle cells proliferated and some of them became large polygonal cells with abundant cytoplasm. When phenyl beta-D-thioxyloside (0.5 mM), an artificial initiator of chondroitin sulfate chain synthesis, was added to the culture medium, the cells showed a marked increase in number as compared to controls, with conversion of about 35% of the cells to large size cells. Immunohistochemical staining with monoclonal antibodies showed that the intercellular matrix of adherent cells consists mainly of a large proteglylcan with chondroitin 6-sulfate side chains. By histochemical analysis, the amount of chondroitin sulfate was shown to be greater in the intercellular matrices of xyloside-treated groups than those of control cultures. The amount of chondroitin sulfate in the growth medium of the adherent cells, as measured by uronic acid analysis, was also significantly increased by treatment with phenyl beta-D-thioxyloside compared with controls.

The frequency and histology of hepatic tissue in germ cell tumors.

The frequency of hepatic tissue and its histological characteristics were examined in 516 germ cell tumors. Hepatic tissue was observed in 48 cases (9.3%). The incidence of hepatic tissue was low in tumors of the ovary (5%), high in both retroperitoneal (27%) and sacro-coccygeal (24%) tumors, and low in both mature (0.3%) and immature teratomas (11%). It was usually encountered in infancy, and the frequency was high in both yolk sac tumors (48%) and mixed germ cell tumors (52%). The hepatic tissue found mainly in mature or immature grade 1 teratomas was similar to adult normal human liver tissue (Ha-type). Tissue in areas consisting of some immature somatic elements of a mixed germ cell tumor was similar to embryonic or fetal liver tissue (Hf-type). Many hepatic nests found in a polyembryoma were of both Ha- and Hf-types. The hepatic tissue found in close relation to yolk sac elements showed predominantly hepatocellular carcinoma-like features (HCLS). Immunohistochemically, the cytoplasm of adult liver-type cells was positive for alpha-1-antitrypsin (AAT), human albumin (ALB), and the third (C3) and fourth (C4) components of the complement system. The cytoplasm of fetal liver-type cells showed the same positivity; in addition, these cells were positive for alpha-fetoprotein (AFP) in 25% of the cases. The cytoplasm of hepatic cells of HCLS was positive for AFP, AAT, ALB, C3, and C4. A weakly positive reaction for CEA and CA19-9 was observed in bile duct-like structure in some Hf-type cases.

Production and characterization of monoclonal antibody to dermatan sulfate proteoglycan.

We purified dermatan sulfate proteoglycan (PG) from the capsule of human ovarian fibroma for use as an immunogen. A monoclonal antibody, designated 6B6, was produced which reacts to the intact molecule of dermatan sulfate PG and the chondroitinase AC-treated core molecule on Western-blotted nitrocellulose membrane. Localization of materials showing crossreactivity to this antibody was studied in human tissues by indirect immunohistochemistry. The interstitial elements of almost all tissues examined were positive for the antibody: dermis, submucosal layer of digestive tract, perichondral layer, perivascular connective tissue, perineurium, adventitia of aorta, vessel wall of vein, pleura, and fibrous capsule of kidney and liver. Positive staining was also observed in fibrous elements at post-necrotic foci of cardiac muscle and pancreas, and at atherosclerotic lesions of aorta. The distribution of the antigen, core protein of the dermatan sulfate PG, revealed with 6B6 was compared to that of the dermatan sulfate side chain, which was demonstrated with antibody 9A-2 (Couchman et al.: Nature 307:650, 1984) after treatment with chondroitin sulfate B-lyase. The distribution of both antigens, core protein, and dermatan sulfate side chains showed the same pattern, with minor exceptions. The antibody 6B6 will be a useful tool to study the immunohistochemical localization of dermatan sulfate PG.

Phylogenetic position of yeast-like symbiont of Hamiltonaphis styraci (Homoptera, Aphididae) based on 18S rDNA sequence.

Almost all aphids harbor bacterial intracellular symbionts in mycetocytes. However, some Cerataphidini aphids do not harbor them but possess yeast-like extracellular symbionts in the abdominal hemocoel, suggesting that in a lineage of this group replacement of symbiont occurred from an intracellular bacterium to an extracellular fungus. To investigate the origin of the newly-acquired symbiont, the 18S rDNA sequence of the yeast-like symbiont of Hamiltonaphis styraci was determined by PCR direct sequencing. Molecular phylogenetic analyses indicated that the symbiont belongs to the subphylum Ascomycotina, the class Pyrenomycetes. It was also suggested that the yeast-like symbiont of H. styraci and that of planthoppers are phylogenetically very closely related to each other.

Ovarian gonadoblastoma with dysgerminoma in a woman with two normal children.

An unusually rare case of unilateral gonadoblastoma with dysgerminoma occurring in the ovary of a 28-year-old woman with two normal children is reported.

Characteristics of "embryoid body" in human gonadal germ cell tumors.

The characteristics of so-called "embryoid body" appearing in gonadal germ cell tumors were studied histologically and immunohistochemically on serial sections of three cases (one ovary and two testes). The embryoid bodies were usually observed to be contiguous with immature or mature intestinal ducts, hepatic nests, or epidermal cell nests on serial sections, though they appeared to be isolated in one section. The "amniotic cavity"-like structure of embryoid body was continuous with intestinal duct, and rarely with squamous cell nests, while the "yolk sac" was continuous with hepatic tissue. In these immature or mature structures, differentiation was always found independently of "disc," and portions of "ectoderm" and "endoderm" remained less differentiated in comparison with others. These findings were in contrast with a normal embryo in which immature and/or mature structures are derived from the embryonic disc. The amniotic cavity connected frequently with yolk sac. From the present results, the embryoid body is not considered to be a real or teratomatous embryo, but only a product during a divergent differentiation into intestine and liver from the plastic epithelium, which seems to be derived from an embryonic gut.

IgD monoclonal gammopathy of undetermined significance.

We describe here a case of monoclonal gammopathy of undetermined significance (MGUS) of IgD type. A trace of serum M-protein was noticed on electrophoresis when a Japanese man visited his primary care physician for medical examination. However, he was asymptomatic. Serum immunoelectrophoresis showed IgD lambda M-protein. Bone marrow examination revealed normocellular marrow with 1.5% plasma cells. Renal function, serum calcium level and complete blood cell count were all within normal limits and a systemic bone survey disclosed no lytic lesions. He was followed for 26 months without chemotherapy, and no disease progression to overt myeloma nor development of systemic amyloidosis resulted. This is, to our knowledge, the third case of IgD MGUS and the first one reported from Japan.

Decreased interleukin-6 level in the cerebrospinal fluid of patients with Alzheimer-type dementia.

We examined IL-6 levels in the cerebrospinal fluid (CSF) of patients clinically diagnosed with Alzheimer-type dementia (ATD) and with vascular dementia (VD) and of age-matched normal subjects. The IL-6 levels in the CSF of ATD, but not VD patients, were significantly decreased. In the early onset ATD patients (disease onset < 65 years), IL-6 levels were reduced to 21% of the control level. The IL-6 levels in the CSF were not associated with the severity of the dementia or the duration of the disease since the identification of the first symptoms.

Solid variant of an aneurysmal bone cyst (giant cell reparative granuloma) of the 3rd lumbar vertebra.

A 9-year-old girl with a solid variant of an aneurysmal bone cyst in the 3rd lumbar vertebra showed a good response to low-dose radiation as the only treatment. The solid variant of aneurysmal bone cyst is thought to be a reactive response to intraosseous hemorrhage and is also called giant cell reparative granuloma or giant cell reaction. These lesions in the jaw and the short tubular bones of the hands and feet frequently recur after surgery. Aneurysmal bone cysts of the spine also show a fairly high recurrence rate after incomplete resection or radiation therapy. However, 7 previous cases of the solid variant of aneurysmal bone cyst in the spine and this case did not show recurrence after a mean follow-up period of 45 months. This difference in behaviour suggests that the solid variant should be recognized before surgery as being distinct from conventional aneurysmal bone cysts, especially in the spine.

Endometrial cytology in postmenopausal hormone replacement therapy.

Endometrial changes is postmenopausal hormone replacement therapy (HRT) were studied by comparing cytological and histological findings. Cytological and histological examinations were conducted on 138 benign cases and 26 abnormal cases, including 24 cases with disordered proliferative phase (DOP) and 2 cases with simple endometrial hyperplasia (SEH), for a total of 164 cases. Hormones were administered as follows: 1) single cyclic administration of estrogen only (Single-HRT) for 31 cases, 2) cyclic administration of estrogen and progestin (Cyclic-HRT) for 105 cases, and 3) continuous administration of estrogen and progestin (Continuous-HRT) for 28 cases. All of the 164 cases were studied cytologically as to shape, appearance, nuclear number on maximum diameter, and so on. The benign cases in each mode of administration as described above revealed the following: 1) Single-HRT, atrophy or the proliferative phase was noted histologically, and the copresence of the endometrial epithelium and the ciliated cell metaplasia was observed cytologically; 2) Cyclic-HRT, the first half of the administration term was of the proliferative phase histologically, and the linear and long glands were seen cytologically. In the latter half of the administration term the secretory phase was noted histologically and the curved/linear glands with subnuclear vacuolization were observed cytologically; and 3) Continuous-HRT, atrophy was noted histologically, and fewer glands and atrophic cells on the endometrial epithelium with wrinkles mixed therein were seen cytologically. On the other hand, cytological examinations of the abnormal cases revealed a mean average of 35 nuclei on the maximum diameter of the gland, protrusion and/or ramification of the glands, densely clustered glands, and back-to-back glands without fusion, as well as irregularly dilated tortuous glands in SEH. These abnormal findings were considered useful for early detection of endometrial disorders in the hormone replacement therapy by cytodiagnosis.

Apocrine carcinoma vs. apocrine metaplasia with atypia of the breast. Use of aspiration biopsy cytology.

OBJECTIVE: To solve the problem of diagnosing apocrine carcinoma (APCA) through distinguishing it from benign apocrine metaplasia with atypia (APMA). STUDY DESIGN: The study group consisted of five histologically confirmed cases of uncommon infiltrating apocrine carcinoma and a case of noninfiltrating apocrine carcinoma of the breast by aspiration biopsy cytology. The control group consisted of 103 cases of benign apocrine metaplasia with no atypia (APMN), 4 cases of APMA and 34 cases of common-type adenocarcinoma that were encountered in 662 breast aspirations from 1988 to 1992 at Hekinan Municipal Hospital. RESULTS: In APCA the average age of patients (65 +/- 17.7 SD)(mean +/- SD) was more than 20 years older than APMA, and APCA generally measured > or = 2 cm or more in diameter as compared to < or = 2 cm in APMA lesions. In APCA the apocrine cells had high cellularity, with the cells occur singly and with syncytia in the background. Numerous degenerated apocrine cells and characteristic cell detritus were found. APCA was also characterized by apocrine cells with more marked nuclear abnormalities, including hyperchromasia and irregular nuclear shape, frequently with irregular nucleoli; more nuclei measure > or = 12 micron in diameter than in APMA. These findings, however, were present only to a mild degree in APMA, if at all. APMA may coexist with APCA. CONCLUSION: If APMA is diagnosed, an open biopsy should be performed to distinguish it from APCA.

True aneurysms of the middle meningeal artery associated with cavernous hemangioma of the skull--case report.

A rare case of true aneurysms of the middle meningeal artery (MMA) associated with cavernous hemangioma in a 47-year-old male is presented. Angiography showed aneurysmal dilatation of the MMA, which was a tumor feeder. Histological examination confirmed the true aneurysmal character of the dilatation. The aneurysms were considered to result from increased hemodynamic stress and medial defects in the MMA.

[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.