Evolutionary discontinuity of the Carabine ground beetles.

The Carabine ground beetles are mostly hind wing-less and cannot fly, so that there is more chance of diversification by geographic isolation compared with winged insects. The relationships between morphological diversification and phylogeny of the ground beetles of the world have been inferred mainly by comparisons of mitochondrial ND5 gene sequences. Based on dating by a mitochondrial DNA "clock," it has been deduced that an explosive radiation of the major carabine groups took place 50-40 MYA. This was followed by occasional radiations on various scales, sometimes accompanied by parallel morphological changes. There are also a good number of examples showing that the fundamental morphology has remained unchanged for a long time among geographically isolated populations within the same species. Thus, carabid evolution would have proceeded discontinuously, with phases of rapid morphological change alternating with silent phases.

Involvement of angiotensin II in progression of renal injury in rats with genetic non-insulin-dependent diabetes mellitus (Wistar fatty rats).

Wistar fatty (WF) rats have a genetic predisposition to hyperglycemia, polyuria, hyperinsulinemia, hyperlipidemia, obesity and nephropathy. These phenotypic characteristics are similar to those observed in obese patients with non-insulin-dependent diabetes mellitus (NIDDM) nephropathy. In this study, the effects of two types of renin-angiotensin system inhibitors, an angiotensin II type 1-receptor antagonist (AT1A) and an angiotensin I-converting enzyme inhibitor (ACEI), on renal injury in WF rats were studied during the progressive phase of diabetic nephropathy. An AT1A, candesartan cilexetil (1 mg/kg), and an ACEI, enalapril (10 mg/kg), were administered orally once a day for 12 weeks, beginning when the rats were 27-week-old and already showed diabetic nephropathy and obesity. Both drugs prevented an increase in proteinuria during the experimental period. Furthermore, after 4-week intervention, the levels of proteinuria were markedly lower in drug-treated rats. At the end of the experiment, both drugs prevented the development of glomerular lesions without affecting glucose metabolism and obesity. In conclusion, the inhibition of angiotensin II activity ameliorated both existing proteinuria and the progression of proteinuria, resulting in preservation of glomerular structure. Thus angiotensin II plays important roles in the development and the progression of nephropathy in genetically obese diabetic WF rats.

Pulmonary multiple metastases of ovarian granulosa cell tumor 15 years after initial diagnosis.

We report a case of multiple pulmonary metastases occurring 15 years after an ovarian granulosa cell tumor was initially diagnosed. A 62-year-old woman undergoing left salpingo-oophorectomy for a granulosa cell tumor of the left ovary 15 years earlier presented with abnormal chest shadows. Computed tomography of the chest confirmed the presence of 3 well-defined nodular lesions, and Computed tomography of the abdomen and pelvis revealed a 3.5 x 2.5 cm partially solid, cystic pelvic mass. Left thoracotomy was conducted and tumors diagnosed a pulmonary metastases of a granulosa cell tumor. The pelvic mass was resected and infracolic omentectomy then conducted with total hysterectomy and right salpingo-oophorectomy including the adherent rectal segment. The pelvic mass proved to be a granulosa cell tumor. Adjuvant combination chemotherapy was started every 3 weeks and the woman has remained disease-free for 9 months.

[Results of surgical treatment for pulmonary aspergilloma in postlobectomy patients].

The purpose of this study was to evaluate the results of surgical treatment of postlobectomy pulmonary aspergilloma. Between January 1986 and September 1999, six patients were surgically treated. Aspergilloma were diagnosed on the basis of the clinical and radiographic findings in all cases. All patients were poor pulmonary function. Space-reducing surgery was performed in 5 patients, completion pneumonectomy was performed in 1 patient. The reoperation was only one, the clinical impression of the cause was too difficult direct closure of the bronchial fistula due to formation of marginal fistula. We believe that space-reducing surgery for postlobectomy patients with pulmonary aspergilloma is useful and benefit treatment.

Formation of the Japanese Carabina fauna inferred from a phylogenetic tree of mitochondrial ND5 gene sequences (Coleoptera, carabidae).

Phylogenetic analyses based on the mitochondrial ND5 gene comparisons and the geohistory of the Japanese Islands suggest that each Japanese species belonging to the subtribe Carabina has its own history for the establishment of its present habitat in the Japanese Islands. It can be roughly classified into two categories: (1) species which were derived from the ancestry that inhabited ancient Japan at the time of its split from the Eurasian Continent [ca. 15 million years ago (MYA)], followed by diversification within the Japanese Islands; and (2) species which invaded Hokkaido from the Eurasian Continent through land-bridges from Sakhalin and/or the Kuriles or from western Japan from the Korean Peninsula during the glacial era (<2 MYA).

Pattern of morphological diversification in the Leptocarabus ground beetles (Coleoptera: Carabidae) as deduced from mitochondrial ND5 gene and nuclear 28S rDNA sequences.

Most of the mitochondrial NADH dehydrogenase subunit 5 (ND5) gene and a part of nuclear 28S ribosomal RNA gene were sequenced for 14 species of ground beetles belonging to the genus Leptocarabus. In both the ND5 and the 28S rDNA phylogenetic trees of Leptocarabus, three major lineages were recognized: (1) L. marcilhaci/L. yokoael/Leptocarabus sp. from China, (2) L. koreanus/L. truncaticollis/L. seishinensis/L. semiopacus/L. canaliculatus/L. kurilensis from the northern Eurasian continent including Korea and Hokkaido, Japan, and (3) all of the Japanese species except L. kurilensis. Clustering of the species in the trees is largely linked to their geographic distribution and does not correlate with morphological characters. The species belonging to different species groups are clustered in the same lineages, and those in the same species group are scattered among the different lineages. One of the possible interpretations of the present results would be that morphological transformations independently took place in the different lineages, sometimes with accompanying parallel morphological evolution, resulting in the occurrence of the morphological species belonging to the same species group (= type) in the different lineages.

Effect of candesartan cilexetil (TCV-116) in rats with chronic renal failure.

BACKGROUND: Inhibition of the renin-angiotensin system by both angiotensin II type 1 receptor antagonists (AT1As) and angiotensin I-converting enzyme inhibitors (ACEIs) shows renoprotective effects in rats with chronic renal failure when treatment is started in the early phase of renal injury. In this study, we examined the renal protective effects of candesartan cilexetil (TCV-116), an AT1A, and enalapril, an ACEI, in the progressive phase of renal injury in 5/6 nephrectomized rats. METHODS: Candesartan cilexetil (1 mg/kg/day) and enalapril (10 mg/kg/day) were orally administered once a day for 4 weeks (the short-term experiment) or 16 weeks (the long-term experiment) to 5/6 nephrectomized rats beginning 15 weeks after the nephrectomy, that is, after they had already showed marked proteinuria. RESULTS: In vehicle-treated rats, proteinuria, glomerulosclerosis, and interstitial fibrosis developed. Moreover, enhanced expression of transforming growth factor-beta1 (TGF-beta1) in the injured glomeruli was observed. These adverse changes progressed with time, and in the short-term experiment, both drugs inhibited them. In the long-term experiment, the progressive proteinuria and the elevation of blood pressure were similarly attenuated by both drugs. However, candesartan cilexetil significantly inhibited the progression of glomerulosclerosis, the expression of TGF-beta1, and interstitial fibrosis, whereas enalapril did not. CONCLUSION: These results indicate that candesartan cilexetil shows potent and long-term preventive effects against the progression of previously developed renal injury.

Evolution of the carabid ground beetles.

The phylogenetic relationships of the carabid ground beetles have been estimated by analysing a large part of the ND5 gene sequences of more than 1,000 specimens consisting of the representative species and geographic races covering most of the genera and subgenera known in the world. From the phylogenetic analyses in conjunction with the mtDNA-based dating, a scenario of the establishment of the present habitats of the respective Japanese carabids has been constructed. The carabid diversification took place ca. 40 MYA as an explosive radiation of the major genera. During evolution, occasional small or single bangs also took place, sometimes accompanied by parallel morphological evolution in phylogenetically remote as well as close lineages. The existence of silent periods, in which few morphological changes took place, has been recognized during evolution. Thus, the carabid evolution is discontinuous, alternatively having a phase of rapid morphological change and a silent phase.

[One case of chronic pyothorax with MRSA infection cured by air-plombage method].

It is very difficult to cure a chronic pyothorax with MRSA infection. We experienced one such case with low pulmonary function (VC 1700 ml, %VC 50.7%), 73 years old man, who had a history of esophageal cancer and was operated two years ago. As the control of bacteria and the surgical intervention are both important in the treatment of pyothorax cases, we tried to reduce MRSA by washing with Povidone-Iodine solution through the drain. Then, we selected Air-plombage method as it is expected to maintain or to increase the pulmonary function after operation. We could easily close the bronchial fistula with a muscle flap, as it was located at the centre of the cavity. During the operation we frequently used acidic electrolyzed NaCl solution against MRSA. For one month after the operation, we used Vancomycin which is effective against MRSA, however, rather severe side effects were seen, and finally and MRSA vanished. Pulmonary function has improved from the initial VC 1700 ml, %VC 50.7% to VC 2120 ml, %VC 63.6% one year later. We recommend Air-plombage method for such cases with low pulmonary function under the control of MRSA by using acidic solution.

Temporal and spatial pattern of expression of the pea phenylalanine ammonia-lyase gene1 promoter in transgenic tobacco.

Genes encoding phenylalanine ammonia-lyase (PAL) form a small multigene family with at least three members in pea. Tissue-specific expression of the promoter of a member of PAL gene family (PSPAL1) was investigated in the transgenic tobacco transformants carrying the different modes of chimeric fusion between the PSPAL1 promoter and a bacterial beta-glucuronidase (GUS) gene. In stems, at least, strict correlation was found between steady-state levels of Gus-mRNA and enzyme activity. Significantly high level of GUS activity was observed in roots, particularly in meristematic tissues and the pigmented region of petals of transgenic tobacco carrying the translational fusion type B (-1,394 to +140 of PSPAL1 connected to Gus), followed by moderately high level of GUS activity carrying the translational fusion type A (-1,394 to +117). GUS expression in tissues of mature leaves, however, was very low in these constructs. Extremely low GUS activity was observed in the transformants of transcriptional fusion type (-1,394 to +5), whilst no activity was detected carrying non-transcription fusion type (-1,394 to -27). Furthermore, the pattern of the PSPAL1 expression was characterized in response to pathogen ingress and woundings in transgenic tobacco carrying the translational fusion type B. Woundings itself triggered marked expression of PSPAL1-driven GUS expression at the wounded sites. Inoculation of nonpathogens, Phytophthora capsici, P. boehmeriae and Erisiphe graminis f. sp. hordei, both caused rapid and very clear GUS expression zone along with the development of hypersensitive cell death area where callose was accumulated, however, the inoculation of a pathogen, P.nicotiana caused slow and hazy GUS expression zone along with the lesion development. These results suggest that the expression of pea PSPAL1 promoter is regulated in a similar fashion, at least in a part, in pea and transgenic tobacco, under the plant development and various environmental cues.

Time-course ultrasonographic observation of a mesenteric pseudocyst of the sigmoid colon: report of a case.

A 35-year-old woman was referred to our hospital for investigation of lower abdominal pain and a feeling of fullness. At her first consultation, a transvaginal ultrasonography (US) revealed a homogeneous cystic mass in the lower abdomen. Over a period of 8 months the US findings of the content of this mass changed from fine and faint internal echoes to moderate amounts of irregularly contoured internal echoes. At laparotomy, the cystic mass, which measured 3.0 x 3.0 x 3.5 cm, appeared to arise in the sigmoid mesocolon and tightly adhered to the appendix. The cyst was unilocular and contained a slightly yellow gelatinous fluid. Microscopically, its wall was fibrous and lacked an epithelial lining, suggesting that it was a so-called pseudocyst arising from the sigmoid mesocolon. To our knowledge, this is the first case report documenting the time-course ultrasonographic observations of a mesenteric pseudocyst. Our findings suggest that the time-elapsed ultrasonographic changes might have been dependent on the interval between the onset of cystic formation and the US examination.

Effects of candesartan cilexetil (TCV-116) and enalapril in 5/6 nephrectomized rats.

The renal protective properties of candesartan cilexetil (TCV-116), an angiotensin II type 1 receptor antagonist (AT1A), and enalapril, an angiotensin I converting enzyme inhibitor (ACEI), were investigated in 5/6 nephrectomized (NX) rats. Candesartan cilexetil (1 mg/kg/day) and enalapril (10 mg/kg/day) were administered orally to 5/6 NX rats for four weeks (during the early phase of disease development) or 16 weeks (through the late phase). In vehicle-treated rats, proteinuria, glomerulosclerosis, interstitial mononuclear cell (MNC) infiltration and interstitial fibrosis developed. Moreover, immunohistological studies showed enhanced expression of transforming growth factor-beta 1 (TGF-beta 1) in the injured glomeruli. Both drugs inhibited these adverse changes in the early phase. In the late phase, the progressive proteinuria, interstitial MNC infiltration were attenuated by both drugs. However, candesartan cilexetil significantly inhibited the progression of glomerulosclerosis, the expression of TGF-beta 1 and the interstitial fibrosis, while enalapril did not. Candesartan cilexetil and enalapril showed comparable hypotensive effects after the 16-week administration. These results indicate that candesartan cilexetil shows a more potent protective effect than enalapril against the progression of renal injury in the late phase. Thus, an AT1A might be more useful than an ACEI for the treatment of patients with chronic renal failure.

[Video-assisted thoracic surgical (VATS) biopsy as a technique for differential diagnosis of indeterminate small pulmonary nodules].

The authors experienced two cases of multiple pulmonary tuberculomas, which were suspected of metastatic lung tumors. First patient was a 63-years-old male, who was found to have multiple pulmonary nodular shadows on his check up chest X-ray film, suspected of pulmonary metastases of rectal cancer. VATS biopsy, performed for one of nodules revealed that it was caseating granuloma, suggestive of pulmonary tuberculosis. Second patient was a 55-years-old female with rheumatoid arthritis and a chest X-ray film showed multiple pulmonary nodules, which were thought to be metastatic lung tumor from unknown origin. VATS biopsy for a subpleural nodule showed epitheloid granuloma with caseous necrosis of lung, indicating pulmonary tuberculosis. VATS biopsy is, therefore, an useful technique for differential diagnosis of small pulmonary nodules of unknown origin.

Antithrombotic effects of TAK-029, a novel GPIIb/IIIa antagonist, in guinea pigs: comparative studies with ticlopidine, clopidogrel, aspirin, prostaglandin E1 and argatroban.

The antithrombotic and bleeding time (BT)-prolonging effects of TAK-029, a novel glycoprotein IIb/IIIa antagonist, were characterized and compared with those of conventional antithrombotic agents in guinea pigs. TAK-029 potently inhibited the binding of fibrinogen and von Willebrand factor to purified human GPIIb/IIIa with IC50 values of 0.67 +/- 0.03 and 0.33 +/- 0.04 nM; it also inhibited human platelet aggregation induced by various aggregating agents with IC50 values of 29 to 38 nM. The in vitro antiplatelet effect of TAK-029 was potent in humans, guinea pigs and monkeys. When TAK-029 was given p.o. to guinea pigs, severe prolonging of BT (>1800 sec) was not observed with plasma concentrations of TAK-029 that inhibited ex vivo platelet aggregation by < 100%. The p.o. administration of TAK-029, ticlopidine and clopidogrel prolonged BT to the same extent, in parallel with their inhibition of ex vivo platelet aggregation. TAK-029 inhibited ex vivo platelet adhesion and thrombus formation in an arteriovenous shunt model more strongly than ticlopidine, clopidogrel and aspirin at doses causing similar prolongations of BT. In a balloon catheter-induced carotid thrombosis model, i.v. administration of TAK-029 significantly inhibited thrombus formation without prolonging BT. At doses that caused an incomplete antithrombotic effect, PGE1-alpha-cyclodextrin and argatroban produced hypotension and prolongation of BT, respectively. TAK-029 may be effective in patients suffering from arterial thrombotic diseases, which are refractory to these conventional antithrombotic agents.

Inhibition of arachidonic acid induced-aggregation of rabbit platelets with CV-4151 (isbogrel), a selective thromboxane A2 (TXA2) synthase inhibitor: modulation of the antiplatelet action and prostanoid metabolism by rat aortic rings.

In rabbit platelet-rich plasma, a specific TXA2 synthase inhibitor, CV-4151 (isbogrel) alone modestly inhibited the platelet aggregation induced by arachidonic acid (AA); even at 10(-4) M the inhibition was less than 50% (IC40 value: 8.1 x 10(-5) M). Aspirin-treated rat aortic rings alone inhibited the AA-induced platelet aggregation by 7%. However, CV-4151 exhibited a potent antiplatelet action in the presence of the aortic rings and its IC40 value was 8.3 x 10(-8) M. Thus, the aortic rings caused 1000-fold enhancement of the antiplatelet action of CV-4151. Next, we investigated the effect of CV-4151 on the production of two highly biologically active prostanoids, TXA2 and PGI2, and the contribution of these prostanoids to the antiplatelet action of CV-4151. TXA2 and PGI2 were measured by a radioimmunoassay of their stable metabolites, TXB2 and 6-keto-PGF1 alpha, respectively. Under the experimental conditions, CV-4151 simultaneously inhibited TXA2 generation and enhanced PGI2 generation. There was a positive linear relationship (r = 0.975, p < 0.01) between % inhibition of platelet aggregation and PGI2 generation and a significant negative linear relationship (r = 0.994, p < 0.001) between % inhibition of platelet aggregation and TXA2 generation. CV-4151 exhibits a potent antiplatelet action in the presence of PGI2 synthase, which is the in vivo situation, by simultaneously inhibiting TXA2 generation and enhancing PGI2 generation.

Microchimerism in female renal transplant recipients from male donors.

Microchimerism in 23 female renal transplant recipients from male donors was studied using nested polymerase chain reaction (nPCR) and fluorescence in situ hybridization (FISH) to detect Y-chromosome. nPCR was a sensitive and specific assay enabling a detection rate of 1/10(6) male/female cells, compared with a sensitivity of 1/10(2) by standard PCR (sPCR). None of the 23 patients with a male allograft demonstrated Y-chromosome using sPCR. In contrast, 1 3 (56.5%) patients demonstrated Y-chromosome with nPCR. Of 9 patients proven to have microchimerism by nPCR, only 3 also demonstrated Y-chromosome using FISH. The existence of B cells and CD8 cells in donor chimeric cells were proved by separation with Dynabeads class I and class II. Dynamic changes of microchimerism occurred in 4 of 5 patients. Four patients were proven to have microchimerism within a year of transplantation and the microchimerism later disappeared in 3, although the sequential change was variable in individual patients. There was no correlation between microchimerism and patients'clinical factors such as donor-specific blood transfusion, HLA matching, immunosuppression, past history of acute rejection and chronic rejection. The degree of microchimerism in renal transplant recipients was relatively low, and its existence did not seem to be compatible with long-term graft acceptance. However, further studies are required to elucidate the immunological mechanism of microchimerism, and it might be an important clue to immunological tolerance.

Beneficial effect of CV-4151 (Isbogrel), a thromboxane A2 synthase inhibitor, in a rat middle cerebral artery thrombosis model.

Effects of thromboxane A2 (TXA2) synthase inhibitors (CV-4151 and ozagrel) on cerebral thrombosis and cerebral damage were examined in a rat middle cerebral artery (MCA) thrombosis model and their potencies were compared with the conventional antithrombotic agents, aspirin and ticlopidine. CV-4151 significantly inhibited photochemically induced MCA thrombosis by oral (1 and 10 mg/kg) and intravenous (1 mg/kg) administration. Ozagrel (10 mg/kg, p.o.) also inhibited it. The potency of CV-4151 was about 10 times stronger than that of ozagrel, being comparable with the inhibition of blood TXA2 generation. Aspirin (100 mg/kg, p.o.) and ticlopidine (300 mg/kg, p.o.) showed an inhibitory tendency on MCA thrombosis. Twenty-four h after photochemical stimulation, cerebral edema and cerebral infarction were observed, and the lactate content in the brain increased. CV-4151 and ozagrel prevented this edema, and the antiedema effects of the drugs were correlated with the antithrombotic effect on thrombotic MCA occlusion. CV-4151 (10 mg/kg, p.o.), furthermore, significantly reduced the infarct size and inhibited the increase in lactate content. These results indicate that TXA2 synthase inhibitors inhibit cerebral damage by inhibition of MCA occlusion with thrombosis, probably resulting from the inhibition of TXA2 generation, and their effects are superior to those of aspirin and ticlopidine. TXA2 might play an important role in cerebral damage in the MCA thrombosis model. CV-4151 might be a useful drug for the treatment of cerebral thrombosis and for the prevention of cerebral infarction.

Effects of thromboxane A2 synthase inhibitors (CV-4151 and ozagrel), aspirin, and ticlopidine on the thrombosis caused by endothelial cell injury.

The antiplatelet and antithrombotic effects of CV-4151 (isbogrel), a potent selective thromboxane A2 (TXA2) synthase inhibitor, were compared with those of ozagrel (OKY-046), aspirin, and ticlopidine in rats. Two hours after oral administration, CV-4151, ozagrel and aspirin inhibited blood TXA2 generation with ID50 values of 0.04, 0.3 and 6.4 mg/kg, respectively. These values were similar to the oral ID50 values of CV-4151 (0.06 mg/kg), ozagrel (0.92 mg/kg) and aspirin (7.0 mg/kg) for arachidonic acid (AA)-induced platelet aggregation ex vivo. Two hours after p.o. administration, CV-4151 and ozagrel inhibited femoral vein platelet-rich thrombosis caused by endothelial injury with ID50 values of 2.46 and 13.7 mg/kg, respectively. However, aspirin (100 mg/kg, p.o.) only slightly inhibited the thrombosis. Ticlopidine (300 mg/kg, p.o.) slightly but significantly inhibited AA-induced and ADP-induced platelet aggregation, however, it potently inhibited the thrombosis. CV-4151 and ozagrel given by i.v. injection showed therapeutic effects on the thrombosis with ED50 values of 0.026 and 0.066 mg/kg, respectively. These values were similar to the i.v. ED50 values of CV-4151 (0.0056 mg/kg) and ozagrel (0.042 mg/kg) for blood TXA2 generation. However, aspirin (30 mg/kg, i.v.) only moderately reduced the thrombosis. CV-4151 (> 0.3 mg/kg, p.o.), ozagrel (> 3 mg/kg, p.o.) and ticlopidine (300 mg/kg, p.o.) all significantly prolonged tail bleeding time. Aspirin (100 mg/kg, p.o.) tended to prolong the bleeding time. The antiplatelet and antithrombotic effects of CV-4151 are more potent than those of ozagrel, aspirin and ticlopidine in rats. CV-4151 may therefore be a useful drug for the treatment of thrombotic diseases.

[Cavernoplasty for atypical mycobacteriosis of the lung--case report of two M. avium complex].

Two patients were diagnosed as suffering from localized atypical myocobacteriosis of the lung consisting of the infection in tuberculous open healing cavity of left upper lobe and giant bulla of right middle lobe. The type of bacteria classified according to Runyon's criteria was Group 3 (M. avium complex). The bacilli had the resistance to most antituberculotics. One of them had the history of pulmonary tuberculosis complicated with giant cavity in left upper lobe and extensive hard pleural adhesion associated with restrictive ventilatory impairment. Another one had the history or right upper lobe lobectomy for tuberculosis complicated extensive pleural adhesion. A combination of thoracoplasty and cavernoplasty was applied to these two cases whose operation were once to the former and twice to the latter respectively, with satisfactory results. The patients show complete closure of the cavity and bulla and have no symptoms of recurrence after surgery. The application of thoracoplasty and cavernoplasty is useful and safe to the treatment of patients suffering from atypical mycobacteriosis of the lung in cases of prospected tough resection, with expecting one stage cure.