Showing papers by "Hokkaido University published in 1999"

Stereochemical Determination of Acyclic Structures Based on Carbon−Proton Spin-Coupling Constants. A Method of Configuration Analysis for Natural Products

Abstract: A method for elucidating the relative configuration of acyclic organic compounds was developed on the basis of carbon-proton spin-coupling constants ((2,3)J(C,H)) and interproton spin-coupling constants ((3)J(H,H)). This method is based on the theory that, in acyclic systems, the conformation of adjacent asymmetric centers is represented by staggered rotamers, and their relative stereochemistry can be determined using (2,3)J(C,H) and (3)J(H,H), because the combined use of these J values enables the identification of the predominant staggered rotamer(s) out of the six possible derived from threo and erythro configurations. Detailed conformational analysis for model compounds 1-4 revealed that this method is useful in most cases for assignment of the configuration of acyclic structures occurring in natural products, in which stereogenic methine carbons are often substituted with a methyl or a hydroxy (alkoxy) group. This J-based configuration analysis was applied to the stereochemical elucidation of carboxylic acid 5 derived from zooxanthellatoxin and proven to be a practical method even for natural products with complicated structures.

The Natural Killer T (NKT) Cell Ligand α-Galactosylceramide Demonstrates Its Immunopotentiating Effect by Inducing Interleukin (IL)-12 Production by Dendritic Cells and IL-12 Receptor Expression on NKT Cells

Abstract: The natural killer T (NKT) cell ligand α-galactosylceramide (α-GalCer) exhibits profound antitumor activities in vivo that resemble interleukin (IL)-12–mediated antitumor activities. Because of these similarities between the activities of α-GalCer and IL-12, we investigated the involvement of IL-12 in the activation of NKT cells by α-GalCer. We first established, using purified subsets of various lymphocyte populations, that α-GalCer selectively activates NKT cells for production of interferon (IFN)-γ. Production of IFN-γ by NKT cells in response to α-GalCer required IL-12 produced by dendritic cells (DCs) and direct contact between NKT cells and DCs through CD40/CD40 ligand interactions. Moreover, α-GalCer strongly induced the expression of IL-12 receptor on NKT cells from wild-type but not CD1−/− or Vα14−/− mice. This effect of α-GalCer required the production of IFN-γ by NKT cells and production of IL-12 by DCs. Finally, we showed that treatment of mice with suboptimal doses of α-GalCer together with suboptimal doses of IL-12 resulted in strongly enhanced natural killing activity and IFN-γ production. Collectively, these findings indicate an important role for DC-produced IL-12 in the activation of NKT cells by α-GalCer and suggest that NKT cells may be able to condition DCs for subsequent immune responses. Our results also suggest a novel approach for immunotherapy of cancer.

In Situ Analysis of Nitrifying Biofilms as Determined by In Situ Hybridization and the Use of Microelectrodes

Abstract: We investigated the in situ spatial organization of ammonia-oxidizing and nitrite-oxidizing bacteria in domestic wastewater biofilms and autotrophic nitrifying biofilms by using microsensors and fluorescent in situ hybridization (FISH) performed with 16S rRNA-targeted oligonucleotide probes. The combination of these techniques made it possible to relate in situ microbial activity directly to the occurrence of nitrifying bacterial populations. In situ hybridization revealed that bacteria belonging to the genus Nitrosomonas were the numerically dominant ammonia-oxidizing bacteria in both types of biofilms. Bacteria belonging to the genus Nitrobacter were not detected; instead, Nitrospira-like bacteria were the main nitrite-oxidizing bacteria in both types of biofilms. Nitrospira-like cells formed irregularly shaped aggregates consisting of small microcolonies, which clustered around the clusters of ammonia oxidizers. Whereas most of the ammonia-oxidizing bacteria were present throughout the biofilms, the nitrite-oxidizing bacteria were restricted to the active nitrite-oxidizing zones, which were in the inner parts of the biofilms. Microelectrode measurements showed that the active ammonia-oxidizing zone was located in the outer part of a biofilm, whereas the active nitrite-oxidizing zone was located just below the ammonia-oxidizing zone and overlapped the location of nitrite-oxidizing bacteria, as determined by FISH.

Terrestrial–aquatic linkages: riparian arthropod inputs alter trophic cascades in a stream food web

Abstract: Dynamics of headwater stream ecosystems are generally regarded as occurring at the interface of aquatic and terrestrial ecosystems Terrestrial arthropod inputs can provide an energy subsidy and increase the abundance of predatory fish, and the ensuing effects potentially can cascade through the food web and ultimately affect primary producers Nevertheless, the community-based effects of such inputs on stream food web dynamics are still poorly understood We present experimental evidence that terrestrial arthropod inputs have an indirect but prominent effect on a stream benthic community by altering the intensity of fish predation in the food web Two key elements of the stream food web, terrestrial arthropod inputs and the presence of predatory fish, were experimentally manipulated by using greenhouse-type covers and enclosures (or exclosures) in a forest stream located in northern Japan When terrestrial arthropod inputs to the stream were experimentally reduced, fish predation pressure shifted dramatically from terrestrial to aquatic arthropods The ensuing depletion of aquatic arthropods resulted in a subsequent increase in periphyton biomass This field experiment revealed that terrestrial arthropod inputs were a primary factor controlling cascading trophic interactions among predatory fish, herbivorous aquatic arthropods, and benthic periphyton These results provide empirical support for the perspective that transfers of energy and biomass from donor systems are frequently significant for the maintenance of biotic communities in recipient systems

An evolutionary programming solution to the unit commitment problem

Abstract: The work was conducted with the aim of finding a general method for solving the unit commitment (UC) problem. The proposed algorithm employs the evolutionary programming (EP) technique in which populations of contending solutions are evolved through random changes, competition, and selection. In the subject algorithm an overall UC schedule is coded as a string of symbols and viewed as a candidate for reproduction. Initial populations of such candidates are randomly produced to form the basis of subsequent generations. The practical implementation of this procedure yielded satisfactory results when the EP-based algorithm was tested on a reported UC problem previously addressed by some existing techniques such as Lagrange relaxation (LR), dynamic programming (DP), and genetic algorithms (GAs). Numerical results for systems of up to 100 units are given and commented on.

Long-term Durability of Dentin Bonds Made with a Self-etching Primer, in vivo

Abstract: The long-term durability of bonds between adhesive resins and dentin is of significant importance for the longevity of bonded restorations. We carried out an in vivo one-year study to evaluate the durability of resin-dentin bonds in the oral cavity, as well as to test the hypothesis that the adhesive interface would show morphological changes in vivo over time. Very shallow saucer-shaped dentin cavities were prepared in 12 intact teeth of one Japanese monkey (Macaca fuscata) under general anesthesia. The cavities were restored with Clearfil Liner Bond II and Clearfil Photo Posterior resin composite. The teeth were extracted at three different times: immediately, and 180 and 360 days after placement of the restorations. One day after the monkey was killed, specimens of the three time periods were subjected to the micro-tensile bond test at a crosshead speed of 1 mm/min. The surfaces of the failed bonds were observed under a field emission scanning electron microscope (FE-SEM). Bond strength measurements in this study were successfully performed and were stable at approximately 19 MPa during the one-year testing. Scanning electron microscopic observations of the failed surfaces revealed, at the top of the hybrid layer and within the adhesive resin, porosity which increased over time. Long-term bonds can be assessed in vivo by the combined evaluation of the microtensile bond strength and SEM morphological examination of the adhesive interface.

Is there a “continental shelf pump” for the absorption of atmospheric CO2?

Abstract: Based on the results obtained in the East China Sea, we propose a new term, Continental Shelf Pump , as a mechanism for the absorption of atmospheric CO 2 . We investigated the carbonate system of the East China Sea along a single observation line traversing its central part on 5 cruises in various seasons. The directly observed fugacity of CO 2 dissolved in the surface water decreased with decreasing salinity and temperature as well as nutrient content. The relation has been expressed as a simple equation of these 3 parameters. Putting the observed data on the parameters in the various parts of the East China Sea in various months into this equation, we have obtained 55 ± 5 ppm as an annual mean fugacity deficit of CO 2 in the surface water of the East China Sea, which nearly equals the directly measured mean fugacity along the observation line. The net absorption flux estimated from the fugacity deficit has agreed with the amount of carbonate transported out of the East China Sea calculated for the distributions of total dissolved carbonate and alkalinity. The distributions of density and total dissolved carbonate reveal the cause of this large deficiency, described as follows. The shallower shelf zone is more cooled than the open sea when heat is lost from the surface. This cooling produces denser water, which together with photosynthetic activity, accelerates the absorption of CO 2 in the shelf zone. The absorbed CO 2 is transformed to organic carbon and regenerated especially at the shallow bottom. Isopycnal mixing (advection and diffusion) transports the denser coastal water, especially the bottom water enriched in dissolved and particulate carbon, into the subsurface layer of the open oceans. The transport continues in the layer below the pycnocline even in the warm season and maintains the low fugacity of CO 2 in the surface water of the shelf zone. This is the continental shelf pump. The pump would account for a net oceanic uptake of CO 2 of 1 GtC/ yr, if the world continental shelf zone would absorb the atmospheric CO 2 at the rate observed in the East China Sea. DOI: 10.1034/j.1600-0889.1999.t01-2-00010.x

Spectroscopic polarimetry with a channeled spectrum.

Abstract: We describe a novel method for the spectroscopic measurement of the state of polarization (SOP) of light. A pair of thick birefringent retarders is incorporated into the spectroscopic polarimeter, so the generated channeled spectrum is composed of three quasi-cosinusoidal components carrying the information about the SOP of the light that is being measured. Fourier inversion of the channeled spectrum provides significant parameters for determination of the spectrally resolved Stokes parameters of light. No mechanically movable components for polarization control or active devices for polarization modulation are used, and all the Stokes parameters can be determined at once from only the single spectrum. The effectiveness of this method is demonstrated by the generation of elliptically polarized light whose SOP varies with wave number.

Resonance in bidecadal and pentadecadal climate oscillations over the North Pacific : Role in climatic regime shifts

Abstract: The roles of interdecadal oscillations in climatic regime shifts, which are observed as rapid strength changes in the Aleutian low in winter and spring seasons, have been analyzed. A regime shift results from simultaneous phase reversals between pentadecadal and bidecadal variations, which synchronize with one another at a relative period of three. The pentadecadal variation, which is observed in both winter and spring seasons, provides the basic timescale of regime shifts, while the bidecadal variation, which is observed only in winter, characterizes the rapidity of the shifts. A Monte-Carlo simulation has shown that the simultaneous phase reversals or resonance between the pentadecadal and bidecadal variations reflect a physical linkage between them and do not coincide accidentally. The role of this synchronization feature for assessing and predicting regime shifts is discussed.

Genetic polymorphism in the 5'-flanking region of human CYP1A2 gene: effect on the CYP1A2 inducibility in humans.

Abstract: A genetic polymorphism was identified in the 5'-flanking region of human CYP1A2 gene, and its effect on the transcriptional activation of the CYP1A2 gene was investigated. Nucleotide sequence analysis revealed the existence of a point mutation from guanine (wild type) to adenine (mutated type) at position -2964 in the gene. This point mutation was detected by a polymerase chain reaction-restriction fragment length polymorphism method using DdeI or BslI restriction enzyme, and was proven to be genetically inherited. Allele frequency in 116 Japanese subjects showed 0.77 and 0.23 for the wild and mutated types of allele, respectively. The point mutation caused a significant decrease of CYP1A2 activity measured by the rate of caffeine 3-demethylation in Japanese smokers (p<0.05). Gel retardation analysis showed the existence of protein bound to the polymorphic locus. These results suggest that this polymorphism is a causal factor of decreased CYP1A2 inducibility.

Incidence and distribution pattern of pelvic and paraaortic lymph node metastasis in patients with Stages IB, IIA, and IIB cervical carcinoma treated with radical hysterectomy.

Abstract: BACKGROUND The incidence and distribution pattern of retroperitoneal lymph node metastasis in patients with cervical carcinoma should be investigated based on data from systematic pelvic lymph node (PLN) and paraaortic lymph node (PAN) dissection, so that a basis can be established for determining the site of selective lymph node dissection or sampling. METHODS A total of 208 patients with Stages IB, IIA, and IIB cervical carcinoma who underwent radical hysterectomy and systematic pelvic and PAN dissection were investigated for lymph node metastasis and histopathologic risk factors for lymph node metastasis. RESULTS Fifty-three patients (25.5%) had lymph node metastasis. The obturator lymph nodes were most frequently involved, with a rate of 18.8% (39/208). Forty-nine of 53 node-positive patients had lymph node metastasis in the obturator, internal iliac, or common iliac lymph nodes. Of 26 solitary lymph node metastases confined to one node group, 18 were in the obturator, 3 in the internal iliac, 3 in the parametrial, and 2 in the common iliac lymph nodes. A multiple logistic regression analysis revealed that deep cervical stromal invasion and lymph-vascular space invasion were related to PLN metastasis. It was also shown that metastasis to bilateral PLNs (excluding the common iliac lymph nodes) as well as metastasis to the common iliac lymph nodes were significantly related to PAN metastasis. CONCLUSIONS The results of this study suggest that the obturator lymph nodes can be sentinel lymph nodes of cervical carcinoma. PAN metastasis appears to occur secondarily to wide-spread PLN metastasis. These results provide a basis for determining the site of selective lymph node dissection and for estimating the existence of PAN metastasis from the pattern of metastasis in PLN in patients with cervical carcinoma. Cancer 1999;85:1547–54. © 1999 American Cancer Society.

Structure, composition and species diversity in an altitude-substrate matrix of rain forest tree communities on Mount Kinabalu, Borneo

Abstract: We studied forest structure, composition and tree species diversity of eight plots in an environmental matrix of four altitudes (700, 1700, 2700 and 3100 m) and two types of geological substrates (ultrabasic and non-ultrabasic rocks) on Mount Kinabalu, Borneo On both substrate series, forest stature, mean leaf area and tree species diversity (both ≥ 48 cm and ≥ 10 cm diameter at breast height [dbh]) decreased with altitude The two forests on the different substrate series were similar at 700 m in structure, generic and familial composition and tree species diversity, but became dissimilar with increasing altitude The decline in stature with altitude was steeper on the ultrabasic substrates than on the non-ultrabasic substrates, and tree species diversity was generally lower on ultrabasic substrates than on non-ultrabasic substrates at ≥ 1700 m The forests on non-ultrabasic substrates at higher altitudes and those on ultrabasic substrates at the lower altitudes were similar in dbh versus tree height allometry, mean leaf area, and generic and familial composition at ≥ 1700 m These contrasting patterns in forest structure and composition between the two substrate series suggested that altitudinal change was compressed on the ultrabasic substrates compared to the non-ultrabasic substrates Tree species diversity was correlated with maximum tree height and estimated aboveground biomass, but was not with basal area, among the eight study sites We suggest that forests with higher tree species diversity are characterized by greater biomass allocation to height growth relative to trunk diameter growth under more productive environment than forests with lower tree species diversity

Tropical climates at the Last Glacial Maximum: a new synthesis of terrestrial palaeoclimate data. I. Vegetation, lake-levels and geochemistry

Abstract: Palaeodata in synthesis form are needed as benchmarks for the Palaeoclimate Modelling Intercomparison Project (PMIP). Advances since the last synthesis of terrestrial palaeodata from the last glacial maximum (LGM) call for a new evaluation, especially of data from the tropics. Here pollen, plant-macrofossil, lake-level, noble gas (from groundwater) and δ18O (from speleothems) data are compiled for 18±2 ka (14C), 32 °N–33 °S. The reliability of the data was evaluated using explicit criteria and some types of data were re-analysed using consistent methods in order to derive a set of mutually consistent palaeoclimate estimates of mean temperature of the coldest month (MTCO), mean annual temperature (MAT), plant available moisture (PAM) and runoff (P-E). Cold-month temperature (MAT) anomalies from plant data range from −1 to −2 K near sea level in Indonesia and the S Pacific, through −6 to −8 K at many high-elevation sites to −8 to −15 K in S China and the SE USA. MAT anomalies from groundwater or speleothems seem more uniform (−4 to −6 K), but the data are as yet sparse; a clear divergence between MAT and cold-month estimates from the same region is seen only in the SE USA, where cold-air advection is expected to have enhanced cooling in winter. Regression of all cold-month anomalies against site elevation yielded an estimated average cooling of −2.5 to −3 K at modern sea level, increasing to ≈−6 K by 3000 m. However, Neotropical sites showed larger than the average sea-level cooling (−5 to −6 K) and a non-significant elevation effect, whereas W and S Pacific sites showed much less sea-level cooling (−1 K) and a stronger elevation effect. These findings support the inference that tropical sea-surface temperatures (SSTs) were lower than the CLIMAP estimates, but they limit the plausible average tropical sea-surface cooling, and they support the existence of CLIMAP-like geographic patterns in SST anomalies. Trends of PAM and lake levels indicate wet LGM conditions in the W USA, and at the highest elevations, with generally dry conditions elsewhere. These results suggest a colder-than-present ocean surface producing a weaker hydrological cycle, more arid continents, and arguably steeper-than-present terrestrial lapse rates. Such linkages are supported by recent observations on freezing-level height and tropical SSTs; moreover, simulations of “greenhouse” and LGM climates point to several possible feedback processes by which low-level temperature anomalies might be amplified aloft.

Apo E structure determines VLDL clearance and atherosclerosis risk in mice

Abstract: We have generated mice expressing the human apo E4 isoform in place of the endogenous murine apo E protein and have compared them with mice expressing the human apo E3 isoform. Plasma lipid and apolipoprotein levels in the mice expressing only the apo E4 isoform (4/4) did not differ significantly from those in mice with the apo E3 isoform (3/3) on chow and were equally elevated in response to increased lipid and cholesterol in their diet. However, on all diets tested, the 4/4 mice had approximately twice the amount of cholesterol, apo E, and apo B-48 in their VLDL as did 3/3 mice. The 4/4 VLDL competed with human LDL for binding to the human LDL receptor slightly better than 3/3 VLDL, but the VLDL clearance rate in 4/4 mice was half that in 3/3 mice. On an atherogenic diet, there was a trend toward greater atherosclerotic plaque size in 4/4 mice compared with 3/3 mice. These data, together with our earlier observations in wild-type and human APOE*2-replacement mice, demonstrate a direct and highly significant correlation between VLDL clearance rate and mean atherosclerotic plaque size. Therefore, differences solely in apo E protein structure are sufficient to cause alterations in VLDL residence time and atherosclerosis risk in mice.

CD44 Variants but not CD44s Cooperate with β1-containing Integrins to Permit Cells to Bind to Osteopontin Independently of Arginine-glycine-aspartic Acid, thereby Stimulating Cell Motility and Chemotaxis

Abstract: The expression of osteopontin (OPN), CD44 variants, and integrins has been correlated with tumorigenesis and metastasis. Here we show that these proteins cooperate to enhance cell motility. First, we demonstrate that several different CD44 variants bind to OPN in an arginine-glycineaspartic acid-independent manner, but that the standard form of CD44 does not. These CD44 variants bind to both the amino- and COOH-terminal portions of OPN independently of the arginine-glycine-aspartic acid sequence, suggesting that multiple domains on OPN can be bound by the CD44 variants. Antibodies directed against the integrin beta1 subunit are able to inhibit this binding. The binding of CD44 variants to OPN is significantly augmented by both anti-CD44s and anti-CD44v antibodies. This augmentation by anti-CD44 antibodies is OPN specific and, again, can be blocked by anti-beta1 antibodies. Finally, we show that OPN binding by CD44 variants/beta1-containing integrins promotes cell spreading, motility, and chemotactic behavior.

Truncated CBP Protein Leads to Classical Rubinstein—Taybi Syndrome Phenotypes in Mice: Implications for a Dominant-Negative Mechanism

Abstract: A mouse model of Rubinstein-Taybi syndrome (RTS) was generated by an insertional mutation into the cyclic AMP response element-binding protein (CREB)-binding protein (CBP) gene. Heterozygous CBP-deficient mice, which had truncated CBP protein (residues 1-1084) containing the CREB-binding domain (residues 462-661), showed clinical features of RTS, such as growth retardation (100%), retarded osseous maturation (100%), hypoplastic maxilla with narrow palate (100%), cardiac anomalies (15%) and skeletal abnormalities (7%). Truncated CBP is considered to have been acting during development as a dominant-negative inhibitor to lead to the phenotypes of RTS in mice. Our studies with step-through-type passive avoidance tests and with fear conditioning test showed that mice were deficient in long-term memory (LTM). In contrast, short-term memory (STM) appeared to be normal. These results implicate a crucial role for CBP in mammalian LTM. Our CBP +/- mice would be an excellent model for the study of the role of CBP in development and memory storage mechanisms.

Posterior occipitocervical reconstruction using cervical pedicle screws and plate-rod systems

Abstract: Study design This retrospective study was conducted to analyze the clinical results in 26 patients with lesions at the craniocervical junction that had been treated by occipitocervical reconstruction using pedicle screws in the cervical spine and occipitocervical rod systems. Objectives To evaluate the effectiveness of pedicle screw fixation in occipitocervical reconstructive surgery and to introduce surgical techniques. Summary of background data Many methods of occipitocervical reconstruction have been reported, but there have been no reports of occipitocervical reconstruction using pedicle screws and occipitocervical rod systems for reduction and fixation. Methods Twenty-six patients with lesions at the craniocervical junction underwent reconstructive surgery using pedicle screws in the cervical spine and occipitocervical rod systems. The occipitocervical lesions were atlantoaxial subluxation associated with basilar invagination, which was caused by rheumatoid arthritis in 19 patients and other disorders in 7. The lowest cervical vertebra of fusion in 16 patients was C2, and the remaining 10 patients underwent fusion downward from C3 to C7. Flexion deformity of the occipitoatlantoaxial complex was corrected by application of extensional force, and upward migration of the odontoid process was reduced by application of combined force of extension and distraction between the occiput and the cervical pedicle screws. Results Solid fusion was achieved in all patients except two with metastatic vertebral tumors who did not receive bone graft for fusion. Correction of malalignment at the craniocervical junction was adequate, and postoperative magnetic resonance imaging showed improvement of anterior compression of the medulla oblongata. There were no neurovascular complications of cervical pedicle screws. Conclusions Occipitocervical reconstruction by the combination of cervical pedicle screws and occipitocervical rod systems provided the high fusion rate and sufficient correction of malalignment in the occipitoatlantoaxial region. Results of this study showed the effectiveness of cervical pedicle screw as a fixation anchor for occipitocervical reconstruction.

Stability and chemical composition of pargasitic amphibole in MORB pyrolite under upper mantle conditions

Abstract: The stability field of pargasitic amphibole in a model mantle composition (MORB pyrolite) has been experimentally determined for a fixed water content. A solidus for a pargasite-bearing lherzolite has been defined at pressures below the limit of amphibole stability of 30 kbar at T = 925 °C. The maximum temperature for pargasitic amphibole in MORB pyrolite occurs at 1075 °C between P = 18 and 25 kbar. This maximum lies between that determined for a fertile peridotite composition (Hawaiian pyrolite) and a depleted peridotite composition (Tinaquillo lherzolite). A comparison of the new results with those from earlier studies suggests that the stability for a particular bulk H2O content is mostly controlled by alkali content of the lherzolite composition. The systematic compositional variation of pargasitic amphibole as a function of pressure and temperature can be represented as an increase of the richterite component with increase in both pressure and temperature. For a given pressure the tschermakite component increases with increasing temperature. The compositions of coexisting clinopyroxenes also show a systematic variation with pressure and temperature. The phase relationships in MORB pyrolite combined with the modal abundance of coexisting phases show that the breakdown reactions of pargasitic amphibole occur continuously throughout the subsolidus region studied. The temperature stability limit of pargasitic amphibole coincides with the water-undersaturated solidus (amphibole-dehydration solidus) at pressures below 30 kbar. The experimental results are applicable to pargasitic amphibole-bearing natural peridotites. Cooling and decompression paths and heating events observed in natural peridotites can be interpreted from changes in the composition of pargasitic amphibole. The data are also applicable to a model for peridotite melting and hydration process in the subduction environment.

Oncogenic Role of Epstein-Barr Virus-Encoded RNAs in Burkitt’s Lymphoma Cell Line Akata

Abstract: Our previous reports indicated that Epstein-Barr virus (EBV) contributes to the malignant phenotype and resistance to apoptosis in Burkitt’s lymphoma (BL) cell line Akata (N Shimizu, A Tanabe-Tochikura, Y Kuroiwa, and K Takada, J Virol 68:6069–6073, 1994; J Komano, M Sugiura, and K Takada, J Virol 72:9150–9156, 1998) Here we report that the EBV-encoded small RNAs (EBERs) are responsible for these phenotypes Transfection of the EBER genes into EBV-negative Akata clones restored the capacity for growth in soft agar, tumorigenicity in SCID mice, resistance to apoptotic inducers, and upregulated expression of bcl-2 oncoprotein that were originally retained in parental EBV-positive Akata cells and lost in EBV-negative subclones This is the first report which provides evidence that virus-encoded RNAs (EBERs) have oncogenic functions in BL cells

Biological Heterogeneity, Including Systemic Replication in Mice, of H5N1 Influenza A Virus Isolates from Humans in Hong Kong

Abstract: An H5N1 avian influenza A virus was transmitted to humans in Hong Kong in 1997. Although the virus causes systemic infection and is highly lethal in chickens because of the susceptibility of the hemagglutinin to furin and PC6 proteases, it is not known whether it also causes systemic infection in humans. The clinical outcomes of infection in Hong Kong residents ranged widely, from mild respiratory disease to multiple organ failure leading to death. Therefore, to understand the pathogenesis of influenza due to these H5N1 isolates, we investigated their virulence in mice. The results identified two distinct groups of viruses: group 1, for which the dose lethal for 50% of mice (MLD 50 ) was between 0.3 and 11 PFU, and group 2, for which the MLD 50 was more than 10 3 PFU. One day after intranasal inoculation of mice with 100 PFU of group 1 viruses, the virus titer in lungs was 10 7 PFU/g or 3 log units higher than that for group 2 viruses. Both types of viruses had replicated to high titers (>10 6 PFU/g) in the lungs by day 3 and maintained these titers through day 6. More importantly, only the group 1 viruses caused systemic infection, replicating in nonrespiratory organs, including the brain. Immunohistochemical analysis demonstrated the replication of a group 1 virus in brain neurons and glial cells and in cardiac myofibers. Phylogenetic analysis of all viral genes showed that both groups of Hong Kong H5N1 viruses had formed a lineage distinct from those of other viruses and that genetic reassortment between H5N1 and H1 or H3 human viruses had not occurred. Since mice and humans harbor both the furin and the PC6 proteases, we suggest that the virulence mechanism responsible for the lethality of influenza viruses in birds also operates in mammalian hosts. The failure of some H5N1 viruses to produce systemic infection in our model indicates that multiple, still-to-be-identified, factors contribute to the severity of H5N1 infection in mammals. In addition, the ability of these viruses to produce systemic infection in mice and the clear differences in pathogenicity among the isolates studied here indicate that this system provides a useful model for studying the pathogenesis of avian influenza virus infection in mammals.

Enantioselective synthesis of binaphthol derivatives by oxidative coupling of naphthol derivatives catalyzed by chiral diamine-copper complexes

Abstract: A highly efficient process of aerobic oxidative coupling of 2-naphthol derivatives catalyzed by 1 mol % of Cu(OH)Cl·TMEDA has been developed. Enantioselective oxidative coupling of naphthols was achieved by the use of 10 mol % of chiral catalysts prepared from proline-derived diamines and cuprous chloride, affording the corresponding binaphthols in good enantioselectivities of up to 78% ee. The ester moiety at the 3-position of the substrate was found to be essential for the good asymmetric induction observed in the present coupling reaction.

Impairment of AMPA receptor function in cerebellar granule cells of ataxic mutant mouse stargazer.

Abstract: The spontaneous recessive mutant mouse stargazer (stg) begins to show ataxia around postnatal day 14 and display a severe impairment in the acquisition of classical eyeblink conditioning in adulthood. These abnormalities have been attributed to the specific reduction in brain-derived neurotrophic factor (BDNF) and the subsequent defect in TrkB receptor signaling in cerebellar granule cells (GCs). In the stg mutant cerebellum, we found that EPSCs at mossy fiber (MF) to GC synapses are devoid of the fast component mediated by AMPA-type glutamate receptors despite the normal slow component mediated by NMDA receptors. The sensitivity of stg mutant GCs to exogenously applied AMPA was greatly reduced, whereas that to NMDA was unchanged. Glutamate release from MF terminals during synaptic transmission to GCs appeared normal. By contrast, AMPA receptor-mediated EPSCs were normal in CA1 pyramidal cells of the stg mutant hippocampus. Thus, postsynaptic AMPA receptor function was selectively impaired in stg mutant GCs, although the transcription of four AMPA receptor subunit genes in the stg GC was comparable to the wild-type GC. We also examined the cerebellum of BDNF knockout mice and found that their MF-GC synapses had a normal AMPA receptor-mediated EPSC component. Thus, the impaired AMPA receptor function in the stg mutant GC is not likely to result from the reduced BDNF-TrkB signaling. These results suggest that the defect in MF to GC synaptic transmission is a major factor that causes the cerebellar dysfunction in the stg mutant mouse.

Effects of Dentin Depth and Cavity Configuration on Bond Strength

Abstract: During polymerization of resin composites, shrinkage stresses compete with resin-dentin bonds in a manner that can cause failure of the bond, depending upon the configuration of the cavity, its depth, and the restorative technique. The hypothesis tested in this study was that the effect of cavity configuration (C) and remaining dentin thickness (RDT) influence resin bond strength to the dentin of Class I cavity floors. The occlusal enamel was ground to expose a flat superficial dentin surface as a control (superficial dentin, C-factor = 1) in human extracted third molars. Cavities 3 mm long x 4 mm wide were prepared to a depth 2 mm below the ground dentin surfaces (deep dentin within cavity floor, C-factor = 3). To assess the relationship between C-factor and RDT, we removed the walls of cavities, making a deep flat surface for bonding (deep dentin, C-factor = 1). The teeth were restored with either Clearfil Liner Bond II (LB II), One-Step (OS), or Super-Bond D Liner (DL), followed by Clearfil Photo Posterior resin composite. After 24 hrs' storage in water, the teeth were sectioned vertically into 3 or 4 slabs (0.7 mm thick) and trimmed for the micro-tensile bond test so that we could determine the strength of the resin bonds to the pulpal floor. All groups gave high bond strengths to superficial dentin, but OS and DL gave significantly lower bond strengths to flat deep dentin when the C-factor was 1. When the C-factor was increased to 3 by the creation of a three-dimensional cavity preparation, the bond strengths of all materials fell (range, 21 to 35%), but the difference was significant (p < 0.05) only with DL. SEM observations of failure patterns showed that specimens with high bond strengths tended to exhibit cohesive failures within the hybrid layer, while specimens exhibiting low bond strengths showed failures at the top of the hybrid layer. Some adhesives do not bond well to deep dentin, making them more susceptible to polymerization shrinkage stress that develops in cavities with high C-factors.

Short-Chain Fatty Acids Suppress Cholesterol Synthesis in Rat Liver and Intestine

Abstract: We previously showed that plasma cholesterol levels decreased following ingestion of a short-chain fatty acid (SCFA) mixture composed of sodium salts of acetic, propionic, and butyric acids simulating cecal fermentation products of sugar-beet fiber (SBF). In the present study, we investigated whether hepatic and small intestinal cholesterol synthesis is involved in the cholesterol-lowering effects of SCFA and SBF. In vitro (expt. 1) and in vivo (expt. 2) cholesterol synthesis rates and the diurnal pattern of SCFA concentrations in portal plasma (expt. 3) were studied in three separate experiments in rats fed diets containing the SCFA mixture, SBF (100 g/kg diet), or the fiber-free control diet. Cholesterol synthesis was measured using 3H2O as a tracer. The in vitro rate of cholesterol synthesis, measured using liver slices, was greater in the SBF group, but not in the SCFA group, than in the fiber-free control group. In contrast, the hepatic cholesterol synthesis rate in vivo was lower in the SCFA group, but not in the SBF group, than in the control group. The mucosal cholesterol synthesis rate for the whole small intestine was <50% of the hepatic rate. The rate in the proximal region was slightly but significantly lower in the SCFA group, and was significantly higher in the SBF group than in the fiber-free group. The rate in the distal small intestines was also significantly greater in the SBF group than in the fiber-free group. Plasma total cholesterol concentrations were lower in the SCFA and SBF groups than in the fiber-free group in both experiments 2 and 3. Diurnal changes in portal SCFA and cholesterol levels were studied in the experiment 3. SCFA concentrations increased rapidly after the start of feeding the SCFA diet, and changes in plasma cholesterol were the reciprocal of those observed in SCFA. These results show that a decrease in hepatic cholesterol synthesis rate mainly contributes to the lowering of plasma cholesterol in rats fed the SCFA mixture diet. Changes in portal SCFA and cholesterol concentrations support this conclusion. In SBF-fed rats, SCFA produced by cecal fermentation are possibly involved in lowering plasma cholesterol levels by negating the counteractive induction of hepatic cholesterol synthesis caused by an increase in bile acid excretion.

Structural changes in intramuscular connective tissue during the fattening of Japanese black cattle: effect of marbling on beef tenderization

Abstract: We investigated changes in structures and mechanical properties of the intramuscular connective tissue during the fattening of Japanese Black steers, using the cell maceration method for scanning electron microscopy. During the early fattening period, from 9 to 20 mo of age, collagen fibrils of the endomysium in longissimus muscle associated more closely with each other, and collagen fibers in the perimysium increased in thickness and their wavy pattern became more regular. These changes were closely related to the increase in mechanical strength of the intramuscular connective tissue and resulted in a toughening of the beef during the period. The shear force value of longissimus muscle decreased after 20 mo of age, concomitantly with the rapid increase in the crude fat content. Scanning electron micrographs of the longissimus muscle dissected from 32-mo-old steers clearly showed that the adipose tissues were formed between muscle fiber bundles, that the honeycomb structure of endomysia was partially broken, and that the perimysium separated into thinner collagen fibers. In semitendinosus muscle, in which the crude fat content was lower (P<.05) than that in longissimus muscle, the structure of the intramuscular connective tissue remained rigid at 32 mo of age. The shear force value of the muscle increased even in the late fattening period, from 20 to 32 mo of age. Thus, the development of adipose tissues in longissimus muscle appears to disorganize the structure of the intramuscular connective tissue and contributes to tenderization of highly marbled beef from Japanese Black cattle during the late fattening period.