Morphological analyses of nephrin expression in progressive glomerulonephropathy of common marmosets

In this study, we focused on nephrin, one of the key molecules within the slit diaphragm of podocytes, as although there have been reports on its expression in humans and rats, their presence in common marmosets has not been reported. We investigated nephrin expression and changes in glomeruli, depending on the development of spontaneous progressive glomerulonephropathy in common marmosets. Nineteen common marmosets at two to ten years of age were evaluated. The kidney was examined by microscopy with hematoxylin and eosin and immunohistochemical staining for nephrin. The lesions were classified into three grades according to a renal lesion grading system reported previously. The nephrin-positive area was measured by morphometric analysis, and the nephrin-positive ratio was calculated. Nephrin expression was observed along the glomerular capillary loop in a continuous linear pattern in renal lesion grades 0 to 2 and either discontinuous linear or coarse granular pattern in grade 3. Nephrin expression tended to decrease significantly depending on the grade of renal lesions. Alteration in nephrin expression has been suggested to play an important role in the progression of renal lesions.     

Microbial metabolites of ingested caffeic acid are absorbed by the monocarboxylic acid transporter (MCT) in intestinal Caco-2 cell monolayers

It was previously reported that m-coumaric acid, m-hydroxyphenylpropionic acid (mHPP), and 3,4-dihydroxyphenylpropionic acid (DHPP) are major metabolites of ingested caffeic acid formed by gut microflora and would be transported by the monocarboxylic acid transporter (MCT). We have directly measured their absorption characteristics in Caco-2 cells using a coulometric detection method involving HPLC-ECD. The proton-coupled directional transport of m-coumaric acid, mHPP, and DHPP was observed, and the transport was inhibited by an MCT substrate. The permeation of m-coumaric acid and mHPP was concentration-dependent and saturable: The Michaelis constant for m-coumaric acid and mHPP was 32.5 and 12.9 mM, respectively, and the maximum velocity for m-coumaric acid and mHPP was 204.3 and 91.2 nmol (min)(-1) (mg protein)(-1), respectively. By contrast, the permeation of DHPP was nonsaturable even at 30 mM and was inversely correlated with the paracellular permeability of Caco-2 cells. Our results demonstrate that these compounds are absorbed by the MCT, although DHPP is mainly permeated across Caco-2 cells via the paracellular pathway. MCT-mediated absorption of phenolic compounds per se and their colonic metabolites would exert significant impact on human health.     

Expression of genes encoding xyloglucan endotransglycosylase/hydrolase in ‘Saijo’ persimmon fruit during softening after deastringency treatment

Persimmon (Diospyros kaki Thunb.) fruit undergoes intensive cell wall modification during postharvest fruit softening. Xyloglucan metabolism is important in cell wall disassembly. We cloned cDNAs for two xyloglucan endotransglycosylase/hydrolase genes (DkXTH1 and DkXTH2) from ‘Saijo’ persimmon fruit treated with dry ice to remove astringency. In order to determine the ethylene dependence of XTH gene expression, fruit were exposed to 1-methylcyclopropene (1-MCP), an inhibitor of ethylene action, prior to removal of astringency. Ethylene production increased in mature control and 1-MCP-pretreated fruit after dry-ice treatment, and flesh firmness decreased to the same extent during dry-ice treatment in the control and 1-MCP-pretreated fruit. After dry-ice treatment, control fruit softened completely, but fruit firmness was maintained in 1-MCP-pretreated fruit. Accumulation of DkXTH1 mRNA was induced simultaneously with commencement of ethylene production in mature control fruit. Pretreatment with 1-MCP delayed accumulation of DkXTH1 mRNA. DkXTH2 expression also coincided with fruit softening but was intensified by 1-MCP treatment during the deastringency treatment. These results indicate that fruit softening was related to both DkXTH1 and DkXTH2 expression in ‘Saijo’ persimmons.     

Greenhouse gas emissions after a prescribed fire in white birch-dwarf bamboo stands in northern Japan,focusing on the role of charcoal

Forest fires affect both carbon (C) and nitrogen (N) cycling in forest ecosystems, and thereby influence the soil–atmosphere exchange of major greenhouse gases (GHGs): carbon dioxide (CO₂), methane (CH₄), and nitrous oxide (N₂O). To determine changes in the soil GHG fluxes following a forest fire, we arranged a low-intensity surface fire in a white birch forest in northern Japan. We established three treatments, having four replications each: a control plot (CON), a burned plot (BURN), and a plot burned with removal of the resulting charcoal (BURN-CHA). Soil GHG fluxes and various properties of the soil were determined on four or five occasions during a period that spanned two growing seasons. We observed increased concentrations of ammonium-N (NH₄-N) in BURN and BURN-CHA after the fire, while nitrate–N (NO₃-N) concentration was only increased in BURN-CHA after the fire. The soil CO₂ flux was significantly higher in CON than in BURN or BURN-CHA, but there was no difference in soil CH₄ uptake between the three treatments. Moreover, the N₂O flux from BURN-CHA soil was slightly greater than in CON or BURN. In BURN-CHA, the soil N₂O flux peaked in August, but there was no peak in BURN. We found temporal correlations between soil GHG fluxes and soil variables, e.g. soil temperature or NO₃-N. Our results suggest that environmental changes following fire, including the increased availability of N and the disappearance of the litter layer, have the potential to change soil GHG fluxes. Fire-produced charcoal could be significant in reducing soil N₂O flux in temperate forests.     

Types of frass produced by the ambrosia beetle <Emphasis Type="Italic">Platypus quercivorus</Emphasis> during gallery construction,and host suitability of five tree species for the beetle

We studied the biology of the ambrosia beetle Platypus quercivorus in the logs of five tree species to determine: (1) the relationship between the amount of frass produced by beetles and tunnel length, (2) the relationship between frass shape and the morphological characteristics of P. quercivorus mouthparts, and (3) the suitability of five tree species for P. quercivorus reproduction. Five logs each from healthy Quercus crispula, Q. serrata, Castanea crenata, Sorbus japonica, and Cryptomeria japonica trees were used in this experiment. The results showed that there was a linear relationship between the amount of frass and tunnel length. Whenever powdery frass was produced, larvae were found in the gallery in the log, while fibrous frass was present only in galleries that contained just adults. The mouthparts of adults were completely sclerotized, which likely accounts for the fibrous frass production. Host preference of P. quercivorus was examined at two stages using five tree species. The first stage is digging initiation, which concerns male preference for digging a tunnel. More holes were made by males on S. japonica and Q. serrata logs, while fewer holes were made on C. japonica logs. The second stage is characterized by female orientation, mating, and progeny development. Platypus quercivorus could complete its lifecycle only in the two Quercus spp.     

Cytosolic-free calcium transients in cultured vascular smooth muscle cells: microfluorometric measurements

Microfluorometric recordings were made of changes in the concentration of cytosolic-free calcium in cultured rat vascular smooth muscle cells treated with quin 2, an intracellularly trapped dye, under several conditions. Nitroglycerin decreased calcium in both the presence and absence of extracellular calcium and strongly and progressively decreased the extent of transient increases in calcium induced by repeated applications of caffeine in the absence of extracellular calcium. Therefore nitroglycerin probably decreases cytosolic-free calcium by accelerating the extrusion of calcium through the sarcolemmal membrane.     

Formation of a one-dimensional array of oxygen in a microporous metal-organic solid

We report the direct observation of dioxygen molecules physisorbed in the nanochannels of a microporous copper coordination polymer by the MEM (maximum entropy method)/Rietveld method, using in situ high-resolution synchrotron x-ray powder diffraction measurements. The obtained MEM electron density revealed that van der Waals dimers of physisorbed O2 locate in the middle of nanochannels and form a one-dimensional ladder structure aligned to the host channel structure. The observed O-O stretching Raman band and magnetic susceptibilities are characteristic of the confined O2 molecules in one-dimensional nanochannels of CPL-1 (coordination polymer 1 with pillared layer structure).     

A three-dimensional synthetic metallic crystal composed of single-component molecules

Molecular metals normally require charge transfer between two different chemical species. We prepared crystals of [Ni(tmdt)2] (tmdt, trimethylenetetrathiafulvalenedithiolate) and carried out crystal structure analyses and resistivity measurements. The analyses and measurements revealed that these single-component molecular crystals are metallic from room temperature down to 0.6 kelvin. Ab initio molecular orbital calculations suggested that pi molecular orbitals form conduction bands. The compact molecular arrangement, intermolecular overlap integrals of the highest occupied and lowest unoccupied molecular orbitals, and tight-binding electronic band structure calculation revealed that [Ni(tmdt)2] is a three-dimensional synthetic metal composed of planar molecules.     

Formation of nanometer-scale grooves in silicon with a scanning tunneling microscope

Grooves a few nanometers wide can be formed on a Si(111) surface with a scanning tunneling microscope when the tip is above a critical voltage. This may provide a promising approach to nanodevice fabrication. The dependence of the critical voltage on tunneling current, tip polarity, and tip material was studied with silver, gold, platinum, and tungsten tips. The results are consistent with field emission of positive and negative silicon ions. The variation of critical voltage with current is explained quantitatively by a simple tunneling equation that includes the effect of the contact potential between tip and sample.