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Similar to psychostimulants, the peripheral administration of menthol promotes mouse motor activity, and the neurotransmitter dopamine has been suggested to be involved in this effect. The present study aimed to elucidate the effects of l-menthol on parts of the central nervous system that are involved in motor effects. The subcutaneous administration of l-menthol significantly increased the number of c-Fos-like immunoreactive nuclei in the dorsal striatum of the mice, and motor activity was promoted. It also increased the extracellular dopamine level in the dorsal striatum of the mice. These observations indicated that after subcutaneous administration, l-menthol enhances dopamine-mediated neurotransmission, and activates neuronal activity in the dorsal striatum, thereby promoting motor activity in mice.

Roman chamomile (Chamaemelum nobile or Anthemis nobilis) has traditionally been used for health care, health maintenance and medicinal purposes, and the essential oil obtained from the flowers is used as a basic ingredient for aromatherapy. Roman chamomile essential oil (CHA) produces psychostimulant-like effects, such as the ambulation-promoting effect, on mouse behaviour. However, mechanisms underlying the effects of CHA remain unclear. The present study aimed to identify the constituents responsible for the ambulation-promoting effect of CHA and to elucidate a possible neuronal mechanism. GC/MS analysis revealed that isobutyl angelate, isoamyl angelate and 2-methylbutyl isobutyrate were the major constituents of CHA. These compounds significantly promoted mouse ambulation, and were thus identified as the major active constituents for the ambulatory effect of CHA. The ambulatory effects were attenuated by chlorpromazine and haloperidol, suggesting that the neurotransmitter dopamine is involved in the ambulation-promoting effects of these constituents similar to already-known psychostimulants. The results suggest that CHA can change a mental condition, at least in part, through the CNS effects of the constituents identified in the current study. The results also suggest that mental effects of CHA are largely different from those of other Roman chamomile preparations due to the composition difference.

Permeability and thermal conductivity of natural gas hydrate sediments were characterized using a microfocus X-ray computed tomography (CT) technique. The free-gas spaces, sand particles, and hydrates or ices were identified from the obtained three-dimensional X-ray CT images, from which continuous pore networks were identified. The absolute permeability of the sediment correlated well increased with the ratio of parallel to vertical directions of continuous pore numbers. This fact indicates that the pore network in sediment can be useful for assessment of permeability. In order to characterize the thermal conductivity of methane hydrate sediments, a homogenization method was applied to microfocus X-ray CT images. The thermal conductivity calculated from three dimensional X-ray CT images was good agreement to those measured by the hot disk method, when the thermal conductivity of sand particles is 0.5 W/mK. These results demonstrated that the microfocus X-ray CT imaging can be effective for studying material flow and thermo-mechanical properties of natural gas hydrate sediments.

The growth of Xe clathrate hydrate in a Xe−ice system was observed in situ by microfocus X-ray computed tomography (CT). In the initial stage of pressurization with Xe gas, quasi-two-dimensional hydrate growth on the ice surface was observed at the Xe−ice interface. With time, Xe hydrate grain grew not only along the ice surface (cover growth), but also toward the gas phase (outer growth), and inside the ice phase (inner growth). A lens-shaped Xe hydrate was formed at the initial interface between the gas phase and ice. Anisotropic hydrate growth resulted in a lens-shaped Xe hydrate. Xe hydrate growth at the gas−ice interface is strongly influenced by the supply of H2O molecules. The inner growth in the lens-shaped Xe hydrate changed in shape from spherical to elliptical. Variations in the contrast in cross-sectional CT images show that the inside of the Xe hydrate region became condensed as time elapsed. Inner growth would result from the formation of a condensed hydrate region, which may lead to a chang...

Our purpose was to clarify factors that influence the level of depression of Japanese children with anorectal malformations (ARM). The subjects comprised 66 children with ARM, aged 0-16 years, and their mothers. Patients were divided into three groups (Group 1: aged 0-5 years, Group 2: 6-11 years, and Group 3: 12-16 years). The level of depression of the children aged 6-16 years was investigated by Kovac's children's depression inventory (CDI). The psychological status of their mothers was assessed by Spielberger's state-trait anxiety index (STAI) and Zung's self-rating depression scale (SDS). The classification of ARM and the clinical condition were also investigated. Defecation scores were assigned for the degree of bowel dysfunction. The level of depression in the patients was more marked in Group 3 than in Group 2 according to the CDI score. Significant correlations between the CDI score of the child and the STAI-1, STAI-2 and SDS scores of the mother were observed in Group 2, but not in Group 3. In Group 2, the STAI-1 score of the mothers was significantly influenced by the degree of bowel dysfunction in her child. In Group 3, the CDI score was significantly correlated with the constipation score. This study revealed that bowel dysfunction is the important factor that influences the level of depression of the children with ARM. In the period of childhood during which the child attends primary school, bowel dysfunction indirectly influences the level of depression of children through the psychological status of mothers. However, bowel dysfunction directly influences the level of depression of children in adolescent patients.

The structure of natural-gas hydrate sediments was studied using a microfocus X-ray computed-tomography (CT) system. The free-gas spaces, sand particles, and hydrates or ices were identified from the obtained three-dimensional (3-D) images. We used CT data to analyze a continuous pore, which allows gas and water flow. The absolute permeability of sediment samples correlated well with horizontal-channel density in terms of direction. The grain-size distribution in sediment samples depended on the spread of flow channels. The average area and length of a channel evidently have little effect on absolute permeability. We determined that absolute permeability increased with the ratio of horizontal- to vertical-channel numbers. It was clear that the number ratio of the horizontal to vertical channels is a predominant factor that determines absolute permeability in similar porosity ranges. These results indicate that the pore network in sediments can be useful for assessing permeability.

Excretory dysfunction associated with congenital anal anomalies (a generic term that includes anal atresia and Hirschsprung's disease) is presumed to greatly affect the psychology of the affected children. In this study, we conducted a survey on the psychological status of children with excretory dysfunction, and investigated the relationship between the affected children and their families in addition to their social life. Four children with fecal dysfunction due to a congenital anal anomaly and their mothers were interviewed. The results of psychological tests in the children and mothers along with our findings in the interviews with the children and mothers, were included in the data analysis. We found that regardless of the degree of excretory dysfunction, the child's state of mind was influenced by whether the mother exhibited warmth or criticism towards her child and whether there was a support system for the mother and child. We suggest that psychiatric consultation is necessary for these children.

We report that various geometric patterns can be formed upon mechanical deformation of hexagonal micro polymer mesh. The patterning of micromesh can be applied to the fabrication of micropatterned soft-materials for cell culturing. A microporous film was prepared from a viscoelastic polymer, poly(e-caprolactone). The film was a hexagonal mesh of 4 μm diameter. Plastic deformation of the film was caused by loading tensile force in one direction. Geometrical patterns such as elongated hexagons, rectangles, squares, and triangles were found in the stretched microporous film. These four types of deformation were reproduced by computer simulations using a viscoelastic network of hexagonally connected viscoelastic bonds. On the stretched hexagonal mesh, cardiac myocytes formed fibrous tissue where cells were aligned along the direction of the long axis of micropores. The hierarchical structure of blood vessels could be modeled by the coculture of endothelial cells and smooth muscle cells using a stretched honey...

A 69-year-old Japanese woman presented with multiple, erythematous, keratotic plaques distributed on her right breast, right shoulder, and left cubital fossa. The lesions varied in size from 5 mm to 6 cm. The patient had first noticed a small area of primary erythema on her left cubital fossa at the age of 18 years. The number and size of the lesions had increased for 50 years. A black nodule appeared on the plaque on the right breast 3 years ago and enlarged to 5 cm in diameter (Fig. 1A). Figure 1. (a) Erythematous plaques varying in size distributed on both breasts. The biggest plaque contained hyperkeratotic tumor. The black nodule appeared on the plaque of the right breast. (b) Six months after black tumor removal, hypertrophic change was noted on the scar. Topical application of vitamin D3 analog caused regression in size and elevation of the largest keratotic plaque, and a decrease in hyperkeratosis of the other eruptions Download figure to PowerPoint The patient was born to healthy nonconsanguineous parents. She grew up in the center of Kanazawa city, Ishikawa prefecture, central area of Japan, and had no history of arsenic exposure or outdoor work. She had no past history of repeated infectious diseases and no family history of recalcitrant warts. A tuberculin test gave an area of induration, 1.0 cm in diameter, in 48 h. The black tumor on the breast was surgically removed with a margin of uninvolved skin. Biopsies from lesions on the right breast (hyperkeratotic erythematous plaque) and left cubital fossa (small erythema without hyperkeratosis) were performed. Microscopic examination of hematoxylin and eosin-stained sections from all specimens showed similar findings. Nests of anaplastic keratinocytes were present in an intraepidermal epithelioma pattern within the expanded epidermal layer. A superficial dermal inflammatory cell infiltrate was also observed. There were irregular masses of epidermal cells that proliferated downwards into the dermis. Atypical mitotic figures, multinucleated keratinocytes, and individual cell keratinization were also observed (Fig. 2). Large clear keratinocytes, specific for epidermodysplasia verruciformis, were not found. Anti-papillomavirus antibody (Dako, Japan) did not react with any of the specimens. A histologic diagnosis of Bowen's disease (BD) (squamous cell carcinoma in situ) was made. Figure 2. In the biopsy specimen from the erythematous plaque on the breast, there were irregular masses of epidermal cells that showed atypical mitotic figures, multinucleation, and individual cell keratinization (hematoxylin and eosin stain, × 100) Download figure to PowerPoint Because the other lesions were distributed diffusely and the patient refused surgical treatment, oral etretinate (10 mg/day) was started; however, dryness of the lips and a burning sensation of the involved skin were intolerable. Therefore, topical application of 1-α,24-dihydroxyvitamin D3 ointment (tacalcitol, Bonalpha™) was chosen as alternative treatment. This vitamin D3 analog provided some relief to the redness and hyperkeratosis of all the erythematous plaques. Moreover, the largest keratotic plaque showed a remarkable regression in size and elevation 6 months later (Fig. 1B). No new tumors or erythematous plaques have been found for 3 years since vitamin D3 analog treatment started. DNA from two frozen tissue samples (black, smooth-surfaced tumor and hyperkeratotic erythematous plaque) was extracted by a standard proteinase K/phenol–chloroform–isoamyl alcohol extraction technique. Samples of total cellular DNA (100 ng) were amplified by polymerase chain reaction (PCR) using four sets of degenerate primers (F10/B5, F12/B5, CP66/CP69, and GP5/GP6) in the highly conserved region of the L1 open reading frame of papillomaviruses (Shamanin V, Delius H, de Villiers EM. Development of a broad spectrum PCR assay for papillomaviruses and its application in screening lung cancer biopsies. J Gen Virol 1994; 75: 1149–1156; Shamanin V, zur Hausen H, Lavergne D, et al. Human papillomavirus infections in nonmelanoma skin cancers from renal transplant recipients and nonimmunosuppressed patients. J Natl Cancer Inst 1996; 88: 802–811; de Villiers EM, Lavergne D, McLaren K, Benton EC. Prevailing papillomavirus types in non-melanoma carcinomas of the skin in renal allograft recipients. Int J Cancer 1997; 73: 356–361). For each set of reactions, negative controls for reagents and DNA were included and processed in the same way as the lesional samples throughout all PCR steps. Two human papillomavirus (HPV) plasmids (0.01 pg) containing the genotypes HPV-4 and HPV-6 served as positive controls. PCR products (10 µL) were subjected to 2% agarose gel electrophoresis. Three sets of primers (F10/B5, F12/B5, and CP66/CP69) amplified HPV-specific DNA fragments in both DNA samples from the BD lesion (Fig. 3). The recombinant plasmids harboring the insert of the amplified DNA were sequenced. The sequences obtained were compared with the available HPV sequences in GenBank (National Center for Biotechnology Information, National Institutes of Health), using the husar software package (Deutsches Krebsforschungszentrum). The sequences of all plasmids corresponded to the L1 portion of HPV-5, except for a few nucleotide substitutions. Figure 3. Results of polymerase chain reaction: 650-bp bands appear in lanes 1, 3, and 4 (human papillomavirus, HPV). Larger sized bands are nonspecific products (NS). Lane M, size marker. Lane 1, DNA extracted from HPV-6 (positive control). Lane 2, DNA extracted from human placenta. Lane 3, DNA extracted from black tumor. Lane 4, DNA extracted from hyperkeratotic plaque Download figure to PowerPoint

We report that tissue-like structure can be formed when cells are cultured on a microporous polymer film (honeycomb film). The honeycomb films were fabricated by applying a moist air to a spread polymer solution containing biodegradable polymers (poly(L-lactic acid) (PLLA) and poly(ε-caprolactone) (PCL)) and an amphiphilic polymer. Hepatocytes were cultured on a self-supporting honeycomb film of PLLA. The hepatocytes formed a single layer of columnar shape cells with a thickness of 20 μm. The tissue formation of hepatocytes was specifically occurred on the honeycomb film of PLLA and not on a flat film of PLLA. Three-dimensional tissue structures were formed, when cells were cultured on both sides of the self-supporting honeycomb film. Double layers of hepatocytes were obtained by the method. Striated tissues such as heart and blood vessel could be reconstructed by utilizing a stretched honeycomb film of PCL.

We report a new fabrication method of a thin polymer film with regular array of micropores (honeycomb film). A honeycomb film was fabricated by depositing a dilute solution of an amphiphilic polymer on water surface. The honeycomb film transferred onto a solid substrate was characterized by atomic force microscopy. By this fabrication method, it was possible to control film area, pore diameter, and film thickness. The pore size depended upon the evaporation time of the polymer solution spread on water surface. The thickness of a honeycomb film was controlled by spreading area of the polymer solution. The spreading behavior was influenced by the water temperature. The film area was proportional to the volume of spread polymer solution and controlled by changing the sub-phase temperature, too. When the polymer solution was simply cast on a solid substrate, a thin polymer layer remained in the bottom of the honeycomb pores. On the other hand, the honeycomb film fabricated on water surface has no bottom layer...

マンガン酸化バクテリアの媒介によってマンガン酸化物が沈殿している東海大学海洋科学博物館の地下海水給水系に, スライド・ガラスを多数設置し, その上に付着するマンガン酸化バクテリアおよび酸化物を時系列的に調べ, マンガン酸化物の成長過程の形態学的および化学的研究を行った。最初, バクテリアがばらばらにスライドガラスに付着し, その後集菌する。それらは集塊状になった後溶菌する。溶菌と共にマンガン酸化物が沈殿を開始し, ドーナツ状, ドーム状を経て, カリフラワー状に成長する。ドーナツ状になった段階で, 酸化物表面に, マンガン酸塩(manganate)特有の蜂の巣状構造が現れる。マンガン酸化物の成長速度は1.8×10-2mmyr-1であり, Mn(II)の酸化速度定数(kMn)は, kMn=6×1019mol-4l4d-1であった。マンガン酸化物中のCoおよびNiの濃度は, 沈殿後取り込み現象(post-depositional uptake)により, 時間と共に増加するが, Cuにはこの傾向は見られなかった。

海水(Mn~2+:約50ppm;pH:約8.0)中でバクテリアの媒介によって生成するマンガン酸化物鉱物を, いろいろなマンガン酸化バクテリアと海水の組み合わせを用いて調べた。その結果, hydrohausmannite, feitknechtite(β-MnOOH), manganite(γ-MnOOH), unnamed MnO2 mineral(JCPDS Card:42-1316)および10Å manganate(buseriteあるいはtodorokite)が生成したが, マンガン酸化物が生成しない組み合わせもかなり存在した。バクテリアのマンガン酸化活性が高いときには, 酸化状態の高いMn(IV)酸化物が, それが低いときには, Mn(III)酸化物が, それぞれ生成した。各鉱物は, 中間物質を経ることなく, Mn~2+の酸化によって直接生成したものと考えられる。海洋環境に存在するMn(IV)酸化物は, 純粋な無機的反応よりもむしろ微生物の媒介によって生成するものと考えられる。

The tensile bond strength (TBS) to denture base materials of silicone-rubber soft relining materials (DRS: Denture Relining Soft type, GC) immersed in water at 37°C from 1 day to 8 weeks was investigated. Four denture base metals (18 K-Au alloy, Ag-Pd-Au alloy, Co-Cr alloy and Pure Ti) and two denture base resins (polycarbonate resin and polysulfon resin) were used. The TBS of DRS was 1.39-2.05 MPa to denture base metals and 1.59-1.81 MPa to denture base resin after 1-dayimmersion, and 1.59-2.75 MPa and 1.47-2.33 MPa respectively after 8-week immersion. These values were significantly superior to those of two ordinary silicone-rubber relining materials.

In order to utilize sarcotestae of Ginkgo biloba for medical purposes, their flavonoids and terpenoids were qualitatively and quantitatively analyzed. Flavonols such as kaempferol, quercetin and isorhamnetin present in leaves could not be detected in sarcotestae while anthocyanidins of delphinidin and cyanidin were found. Sarcotestae were noted to contain small amounts of ginkgolides A, B and C as terpenoids, as did also the leaves. The composition of ginkgolides in sarcotestae were varied during maturation whereas quantity of anthocyanidins was independent on harvest season.

The structure of natural gas hydrate sediments was observed by microfocus X-ray computed tomography (CT). A newly developed high-pressure vessel for the microfocus X-ray CT system was applied to observe the sediments at a temperature above 273 K and under high-pressure conditions. The obtained two-dimensional CT images clearly showed the spatial distribution of the free-gas pore, sand particles, water, and hydrates. These results demonstrated that microfocus X-ray CT can be effective for studying natural gas hydrate sediment samples.

The mammalian target of rapamycin complex 1 (mTORC1: mTOR-raptor interaction) and heat shock protein 70 (Hsp70) regulate various cellular processes and are crucial for the progression of many cancers and metabolic diseases. In the recent study, we reported that interaction of Hsp70 with tuberous sclerosis complex 1 (TSC1) regulated apoptosis. This study was designed to elucidate the underlying mechanism in Cos-1 cells. Here, we show that N-formyl-3,4-methylenedioxy-benzylidene-gamma-butyrolaetam (KNK437), which inhibits the expression level of Hsp70, abrogated phosphorylation of mTOR and S6K in response to insulin, and inhibited mTORC1 activity via disruption of an interaction between mTOR and raptor. In addition, KNK437 did not alter TSC1/2 complex formation. Furthermore, KNK437 inhibited the mTOR-raptor interaction on the outer membrane of the mitochondria and triggered caspase-3 activation. A reduction in the level of Hsp70 could result in the inhibition of the mTORC1 signaling pathway, thereby inducing apoptosis.

To determine whether noradrenergic projections to the posterior hypothalamus via baroreflex are altered in the hypertensive state, the extracellular norepinephrine (NE) content of the posterior hypothalamus was measured in both spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) after sinoaortic denervation (SAD) or sham operation. In WKY, blood pressure (BP) and extracellular NE content 24 h after SAD were significantly higher than those of sham-operated rats. Contrarily, SAD did not increase both BP and NE in the posterior hypothalamus of SHR. These results suggest that the noradrenergic pathway via the baroreflex is impaired in SHR. This mechanism may play an important role in the development and maintenance of hypertension in SHR.

To determine whether chronic central gamma-aminobutyric acid (GABA) stimulation would attenuate development of hypertension, tail-cuff systolic pressures were measured in male spontaneously hypertensive rats (SHR) treated with either an inhibitor of GABA breakdown (valproate, VPA), or a GABA-receptor agonist (muscimol). When VPA was injected intraperitoneally (i.p.) daily for 4 weeks, tail-cuff systolic pressures were decreased during the last 2 weeks. On the other hand, continuous infusion of muscimol into the lateral cerebral ventricle for 14 days using an osmotic minipump decreased systolic pressures throughout the 10-day observation period. During terminal experiments under urethane anesthesia, pressor and sympathetic nerve responses to electrical stimulation of the ventromedial hypothalamus were reduced in VPA- or muscimol-treated SHR. Pressor responses to intravenously (i.v.) injected norepinephrine (NE) were unaltered by either treatment, indicating that diminished pressor responsiveness to hypothalamic stimulation was not due to diminished cardiovascular reactivity. Regardless of either VPA acid or muscimol treatment, basal blood pressure (BP), heart rate (HR), and responses to hypothalamic stimulation were lower in normotensive Wistar-Kyoto controls (WKY) than in SHR. In addition, chronic treatment with either drug had weaker hypotensive and hypothalamic depressant effects in WKY than in SHR. Our results suggest that in SHR the central GABAergic system is impaired and that chronic treatment with GABAergic stimulants can attenuate development of spontaneous hypertension by reducing hypothalamic overactivity.

A 59 year-old housewife was admitted to the emergency service with a sudden onset of chest pain and nausea. Initially she was treated as an acute myocardial infarction, but conventional treatments were not effective, and she was sent to our hospital for further evaluation. Her ECG showed several abnormal findings including T-wave inversion, atrial flutter, QT-time prolongation, ST-segment depression or elevation, and frequent ventricular ectopic beats. The echocardiogram, 201thallium scintigram and coronary angiography were almost normal. Both urinary and plasma levels of catecholamines were remarkably increased, and the plasma epinephrine was extremely high during attacks. Abdominal echotomography and CT-scanning showed a large left adrenal tumor. The 131MIBG scintiscan revealed a high accumulation in this tumor. Then the patient was diagnosed as having pheochromocytoma and catecholamine-induced myocarditis. The administration of phentolamine (10 mg) normalized the inversion of T-wave and the high blood pressure. But when propranolol (2 mg) was administrated in addition to phentolamine, the ECG showed a biphasic low T-wave change. According to these phenomena, we supposed that the alpha-adrenergic receptor was involved in the development of the ST-T changes of the ECG, and the alpha-adrenergic receptor of this patient might be sensitive under excessive catecholamines, according to the inhibition of the beta-receptor by propranolol.

To determine whether clonidine would attenuate exaggerated pressor responses to stress in spontaneously hypertensive rats (SHR), pressor responses to shaker stress were recorded following intracerebroventricular (icv) injections of clonidine in conscious SHR. Pressor responses to shaker stress were significantly larger in SHR than in Wistar-Kyoto rats (WKY). Increased sympathetic nerve responses to shaker stress were also significantly greater in SHR than in WKY. Exaggerated pressor responses in SHR were partly attenuated by adrenalectomy and abolished by clonidine (icv). By contrast, 1-Sar,8-Ile angiotensin II (icv) was ineffective. These findings suggest that in SHR, the alpha-adrenergic system in the brain may contribute to exaggerated pressor responses to shaker stress, but not the brain renin-angiotensin system.

Schizencephalyis a rare disorder of brain development resulting in the formation of abnormal unilateral or bilateral clefts in the cerebral hemispheres. One patient is described with psychotic symptoms with unilateral schizencephaly. The implications of the association between schizencephaly and psychosis in these patients for understanding the biology of the psychoses are discussed.

Objective The aim of this study was to investigate the psychological status of Japanese children with congenital anorectal malformation and their mothers to develop appropriate psychiatric interventions. Method The subjects comprised 50 children with congenital anorectal malformation aged 0 to 16 years and their mothers. The psychology of children aged 7 to 16 years was investigated by Kovacs' Children's Depression Inventory (CDI). The psychology of their mothers was assessed by Spielberger's State-Trait Anxiety Index (STAI) and Zung's Self-rating Depression Scale (SDS). Results Depression was more marked in the children aged 12 to 16 years than in those aged 7 to 11 years according to the CDI. The mothers of preschool children showed higher levels of anxiety and depression than those of school-aged children, according to the STAI and SDS. Significant correlations between the CDI score of the children and the STAI or SDS score of the mothers were observed only among children aged 7 to 11 years. Conclusions The frequency of depression and anxiety among children with congenital anorectal malformation and that among their mothers was associated with the age of the child. Long-term postoperative psychosocial support for the children and their mothers may be required, taking into account the age of the child.

Tohoku University has developed various artificial organs over the last 30 years. Pneumatic driven ventricular assist devices with a silicone ball valve have been designed by the flow visualization method, and clinical trials have been performed in Tohoku University Hospital. On the basis of these developments, a pneumatic driven total artificial heart has been developed and an animal experimental evaluation was conducted. The development of artificial organs in Tohoku University has now progressed to the totally implantable type using the transcutaneous energy transmission system with amorphous fibers for magnetic shielding. Examples of implantable systems include a vibrating flow pump for ventricular assist device, an artificial myocardium by the use of shape memory alloy with Peltier elements, and an artificial sphincter for patients with a stoma. An automatic control system for artificial organs had been developed for the ventricular assist devices including a rotary blood pump to avoid suction and to maintain left and right heart balance. Based upon the technology of automatic control algorithm, a new diagnostic tool for evaluating autonomic nerve function has been developed as a branch of artificial organ research and this new machine has been tested in Tohoku University Hospital. Tohoku University is following a variety of approaches aimed at innovation in artificial organs and medical engineering fields.

Gastrol (1), together with 10 known phenolic compounds, has been isolated from the MeOH extract of the rhizomes of Gastrodia elata Blume (Orchidaceae), and their structures were elucidated by detailed spectral analyses including by 2D NMR spectroscopic analyses. The relaxant effects of these constituents on smooth muscle preparations isolated from guinea-pig ileum were also studied in order to reveal their characteristic pharmacological activities.

We report that microporous films (honeycomb films) can lead various types of cells to tissue formation. The honeycomb films were fabricated by applying a moist air to a spread polymer solution containing biodegradable polymers (poly(L-lactic acid) (PLLA) and poly (ε-caprolactone) (PCL)) and an amphiphilic polymer. Hepatocytes were cultured on a self-supporting honeycomb film of PLLA. The hepatocytes formed a single layer of columnar shape cells with a thickness of 20 μm. The tissue formation of hepatocytes specifically occurred on the honeycomb film of PLLA, not on a flat film of PLLA. Three dimensional tissue structures were formed, when cells were cultured on both sides of the self-supporting honeycomb film. Double layers of hepatocytes were obtained by the method. Striated tissues such as heart and blood vessel could be reconstructed by utilizing a stretched honeycomb film of PCL.

We propose a model of the intracellular delivery process in which fluorescein-labeled natural oligonucleotides (F-DNA) are transferred into the nuclei of cultured human keratinocytes. By encapsulation in neutral multilamellar lecithin liposomes, the F-DNA appeared to be protected against intracellular interactions with cellular materials and nuclease attacks in the cytoplasm during the process. The intracellular behavior of F-DNA and fluorescent phospholipid-labeled liposomes was observed by means of fluorescence analysis. Results showed that: F-DNA encapsulated in neutral multilamellar liposomes reached the cellular nuclei more efficiently than either free F-DNA, or F-DNA in unilamellar liposomes; the liposomal membranes appeared to be left in the cytoplasm. The reaction of F-DNA with complementary DNA was suggested by a rapid quenching of the fluorescence in the nucleus. In addition, the fluorescence decrease was evidently suppressed in the cytoplasm, indicating a protective effect of the neutral multilamellar liposomes against the interaction of F-DNA with cytoplasmic materials. The application of these findings to ‘photo’-antisense studies has been discussed, where suppression of a gene expression is attempted by using oligonucleotide-attached fluorescein with the aid of a photo-induced covalent binding property.

CdSe nanoparticles were synthesized in the cavity of the cage-shaped protein, apoferritin, by designing the slow chemical reaction system of Cd 2 + and Se 2 - . The cavity size of apoferritin molecules is around 7 nm and the diameter of synthesized CdSe nanoparticles is about 6 nm with small size dispersion. CdSe nanoparticles as prepared are multi-crystalline and can be made into a single crystal by the heat-treatment under nitrogen gas. This bio-template synthesis provides a new method to synthesize uniform CdSe nanoparticles.

From biological and clinical perspectives, it is important to clarify tumor heterogeneity. This study was aimed to investigate whether or not intratumoral heterogeneity of DNA ploidy pattern exists in hepatocellular carcinoma (HCC).Using fresh materials resected from 31 untreated patients, DNA ploidy was analyzed at different sites of the same HCC by means of flow cytometry. The tumor size ranged from 1.7 to 25.0 cm.There was no case in which euploid and aneuploid HCCs coexisted in the same tumor. The DNA ploidy pattern was euploid in 15 and aneuploid in 16 instances. Of 15 euploid tumors, the areas analyzed were all diploid in 12 and tetraploid in 2 but diploid/tetraploid in 1. Among 16 aneuploid tumors, the DNA indices (DI) at different sites were similar in 9, but apparently, different aneuploid subclones coexisted in 7 cases. The incidence of DI heterogeneity in aneuploid HCCs was similar between small (5 cm) and large (or = 5 cm) tumors; 3 of 7 (42.9%) versus 4 of 9 (44.4%).It is assumed that euploid and aneuploid HCCs develop in their own ploidy pattern and that the evolution of aneuploid subpopulations from euploid HCC is rare, but new aneuploid subclones can evolve from aneuploid HCC due to increased instability of its karyotype.

To determine whether clonidine can improve the central alteration of the baroreflex in SHR, the centrally cut end of the aortic depressor nerve was electrically stimulated in SHR with intravenously administered clonidine. Aortic depressor nerve (ADN) stimulation elicited depressor and sympatho-inhibitory responses in a frequency-dependent manner in WKY and SHR. These responses were significantly smaller in SHR than in WKY. The attenuated depressor and sympatho-inhibitory responses to ADN stimulation in SHR were restored following i.v. injections of clonidine, although the drug did not affect the responses to ADN stimulations in WKY. These findings suggest that clonidine can improve central attenuation of the baroreflex in hypertension.

This study aimed to elucidate the mechanism of the hypotensive effect of bromocriptine (BRC), and to investigate whether or not the effects of BRC on the sympathetic nervous system are altered in hypertension. BRC was administered intravenously to normotensive and spontaneously hypertensive rats (SHR). It elicited hypotensive effects dose-dependently in urethane-anaesthetized normotensive rats, an effect which was antagonized with metoclopramide. Pretreatment with intravenous hexamethonium attenuated the hypotensive effect of BRC. BRC decreased plasma norepinephrine (NE) without inhibiting the sympathetic nerve spikes recorded from the postganglionic sympathetic nerve bundle. The hypotensive effect of BRC was significantly greater in SHR than in Wistar Kyoto Rats (WKY). Decrease in NE by BRC was also significantly greater in SHR than in WKY. These results suggest that the hypotensive effect of BRC is induced by suppression of NE release, not by inhibition of sympathetic nerve spikes, and that the dopaminergic presynaptic inhibition is attenuated in SHR.

To determine whether the paraventricular nucleus (PVN) contributes to the development of hypertension in spontaneously hypertensive rats (SHR), we compared cardiovascular responses to ganglionic blockade with hexamethonium or vasopressin antagonism with dPVAVP in sham-operated or PVN lesioned SHR and Wistar-Kyoto rats (WKY). Lesions were produced electrolytically when the rats were 5 weeks old. During the next 3 weeks, tail-cuff measurements showed that the development of hypertension in SHR was inhibited, while systolic pressure in WKY was unaffected. Mean pressures recorded directly from the femoral artery at 8 weeks of age were lower in lesioned than in sham-operated SHR (141 +/- 5 vs 110 +/- 3 mm Hg, P less than 0.05), but did not differ in corresponding WKY groups (110 +/- 4 vs 112 +/- 5 mm Hg). Depressor responses to ganglionic blockade induced by i.v. injection of hexamethonium (25 mg/kg) were significantly larger in sham-operated than in lesioned SHR (-41 +/- 4% vs -28 +/- 3%, P less than 0.05). By contrast, vasopressin antagonism with dPVAVP did not alter blood pressure in all rat groups. In 24-h urine samples, excretion of vasopressin was unaffected, but that of norepinephrine was significantly reduced in lesioned SHR. These findings suggest that the PVN contributes to the development of spontaneous hypertension by sympathetic activation without increasing vasopressin secretion.