Ex vivo bupivacaine treatment results in increased adipogenesis of skeletal muscle cells in the rat

Intramuscular adipose tissue (IMAT) is observed in some skeletal muscle pathologies. IMAT is implicated not only in the disorders of muscle contraction, but also of metabolism and insulin sensitivity due to its nature as a secretary organ. Several studies indicate the presence of cells with adipogenic potential in skeletal muscle. However, the mechanism of fate specification that triggers these cells to enter an adipogenic program in vivo remains to be solved. In the present study, we examined whether activation of the adipogenic program of muscle‐resident cells precedes their proliferation upon muscle injury. For this purpose, muscle injury was induced by injecting bupivacaine (BPVC) to excised skeletal muscle ex vivo. Cells isolated from ex vivo BPVC‐treated muscle exhibited higher adipogenic potential than those from saline‐treated muscle. Pre‐plating exposure of skeletal muscle cells to basic fibroblast growth factor (bFGF) mimicked the effect of ex vivo BPVC‐treatment, suggesting that bFGF released from extracellular matrix in response to muscle injury activates their adipogenic program. Interestingly, the number of myotubes were significantly reduced in the culture from BPVC‐treated muscle, suggesting that adipocytes negatively regulate myogenesis.     

Molecular relationships between Australian annual wild rice,Oryza meridionalis,and two related perennial forms

Background

The perennial, Oryza rufipogon distributed from Asia to Australia and the annual O. meridionalis indigenous to Australia are AA genome species in the Oryza. However, recent research has demonstrated that the Australian AA genome perennial populations have maternal genomes more closely related to those of O. meridionalis than to those found in Asian populations of O. rufipogon suggesting that the Australian perennials may represent a new distinct gene pool for rice.

Results

Analysis of an Oryza core collection covering AA genome species from Asia to Oceania revealed that some Oceania perennials had organellar genomes closely related to that of O meridionalis (meridionalis-type). O. rufipogon accessions from New Guinea carried either the meridionalis-type or rufirpogon-type (like O. rufipogon) organellar genomes. Australian perennials carried only the meridionalis-type organellar genomes when accompanied by the rufipogon-type nuclear genome. New accessions were collected to better characterize the Australian perennials, and their life histories (annual or perennial) were confirmed by field observations. All of the material collected carried only meridionalis-type organellar genomes. However, there were two distinct perennial groups. One of them carried an rufipogon-type nuclear genome similar to the Australian O. rufipogon in the core collection and the other carried an meridionalis-type nuclear genome not represented in the existing collection. Morphologically the rufipogon-type shared similarity with Asian O. rufipogon. The meridionalis-type showed some similarities to O. meridionalis such as the short anthers usually characteristic of annual populations. However, the meridionalis-type perennial was readily distinguished from O. meridionalis by the presence of a larger lemma and higher number of spikelets.

Conclusion

Analysis of current accessions clearly indicated that there are two distinct types of Australian perennials. Both of them differed genetically from Asian O. rufipogon. One lineage is closely related to O. meridionalis and another to Asian O. rufipogon. The first was presumed to have evolved by divergence from O. meridionalis becoming differentiated as a perennial species in Australia indicating that it represents a new gene pool. The second, apparently derived from Asian O. rufipogon, possibly arrived in Australia later.     

The coat protein of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis> L<Subscript>11</Subscript>Y is associated with virus-induced chlorosis on infected tobacco plants

The L₁₁Y strain of Tomato mosaic virus (ToMV) causes severe chlorosis on infected tobacco leaves. Sequencing analysis for the genome showed that L₁₁Y contained multiple nucleotide changes and that some led to amino acid substitutions, when compared with that of the common L strain of ToMV. The chimeric virus, which has the CP of L₁₁Y in the context of the L strain RNA genome, caused severe chlorosis on infected tobacco plants, suggesting that the CP of L₁₁Y containing three amino acid changes (E33S, A86T and E97K) was the determinant of the chlorosis. Two of these amino acid changes (A86T and E97K) were associated with the induction of chlorosis when present together in the CP. Severe destruction and deformation of chloroplasts and the formation of discrete dark-staining materials adjacent to chloroplasts were observed with electron microscopy in L₁₁Y-infected plants. Fewer virus particles accumulated in the cytoplasm of L₁₁Y-infected plant cells. The level of accumulation of CP subgenomic RNA and CP in the infected protoplasts was similar between L and L₁₁Y. Fewer virus particles accumulated in L₁₁Y-infected protoplasts, and many of them were shorter-than-full-length. The nucleotide sequence data reported is available in DDBJ/EMBL/GenBank databases as accession AB355139.     

Larval survival,development and growth in the horsehair crab,Erimacrus isenbeckii,cultured under different photoperiod conditions

Larval sinking causes larval mass mortality during seed production in the horsehair crab, Erimacrus isenbeckii. Under normal light conditions, horsehair crab larvae generally show negative phototactic behaviour and sink in their rearing seawater. It has been proposed that culturing larvae in the dark may prevent larval sinking. Herein, we examined the effect of photoperiods on horsehair crab larval survival and development to facilitate the development of larval rearing techniques that prevent sinking. Newly hatched larvae were reared with Artemia to the first crab stage in 2‐L beakers under five photoperiods: 0L:24D, 6L:18D, 12L:12D, 18L:6D and 24L:0D. Larvae survived and molted to the first crab stage under all tested photoperiod conditions. The survival rate improved with increasing light period, whereas the developmental period for each larval stage decreased with increasing light period. Longer light periods increased the carapace length at the first crab stage. Our results suggest that larvae could be cultured to the first crab stage in large‐scale tanks under constant darkness. However, significantly improved larval performance under longer photoperiodic conditions indicates a need for developing alternative culture techniques to control larval behaviour in the seed production tank.     

Effects of NPK fertilizer combinations on yield and nitrogen balance in sorghum or pigeonpea on a vertisol in the semi-arid tropics

A long-term experiment was carried out on a Vertisol from 1986 to 1992 to examine the combined effects of NPK fertilizers on yield using sorghum (Sorghum bicolor L. Moench cv. CSH 5) and short-duration pigeonpea (Cajanus cajan L. Millsp. cv. ICPL 87). The fertilizer treatments were as follows: 0 (no fertilization), N (150 kg N ha^-1 ), P (65.5 kg P₂O₅ ha^-1), K (124.5 kg K₂O ha^-1), and all possible combinations (NP, NK, PK, and NPK). In this study we continued this experiment during the period 1993 to 1994 and analyzed the crop yield response to fertilizers and the N balance. The amount of N derived from the atmosphere and fertilizer was estimated by the ¹⁵N natural abundance method and ^l5N isotope dilution method, respectively. A combined application of Nand P fertilizers gave the highest grain yield for the two crops under the 8th and 9th continuous croppings, unlike the application of K fertilizer. The values of total N for the two crops were significantly higher in the NP and NPK plots. These crops took up N mainly from soil. There was a significant positive relationship between the uptake of N_dff and N_dfs by each crop. Pigeonpea or sorghum took up more N from the soil in the N fertilizer plots than in the plots without N, suggesting that soil N fertility was enhanced and the amount of N supplied from soil increased in the plots with consecutive application of N fertilizer for 7 y. Even pigeonpea, which fixes atmospheric N inherently, needed N fertilizer to achieve high grain yield, suggesting that N fixation by the nodules was not always sufficient to meet the N requirements of the crop under these conditions. Although fertilizer N exerted a beneficial effect on plant growth and yield in the two crops, the values of fertilizer N recovery (FNR) by the two crops were considerably low. Therefore, it is suggested that the development of N fertilizer management which could maximize FNR of each crop should be promoted.     

Antioxidation mechanism studies of caffeic acid: identification of antioxidation products of methyl caffeate from lipid oxidation

As a part of a research project on the elucidation of the chain-breaking antioxidation mechanism of natural phenols in food components, caffeic acid, a polyphenolic acid widely distributed in edible plants, was investigated. The identification and time course analysis of the antioxidation reaction products from methyl caffeate were carried out in the ethyl linoleate oxidation system. The antioxidation reaction produced a quinone derivative of methyl caffeate as an antioxidation product during the initial stage, which was identified by (13)C NMR. The quinone, however, was not the final product, and a further reaction occurred to produce several new peroxides. The isolation and structure determination of the peroxides revealed that they had tricyclic structures, which consisted of ethyl linoleate, methyl caffeate, and molecular oxygen. On the basis of the formation pathway of these products, an antioxidation reaction mechanism of methyl caffeate, including the redox reaction of the caffeate and Diels-Alder reaction of the produced peroxides, was proposed.     

Effects of Artemia enriched with eicosapentaenoic and docosahexaenoic acid on survival and occurrence of molting failure in megalop larvae of the mud crab Scylla serrata

ABSTRACT:   The present study was conducted to investigate the effect of eicosapentaenoic acid (20:5n-3, EPA) and docosahexaenoic acid (22:6n-3, DHA) on the survival and the occurrence of molting failure to megalops of mud crab Scylla serrata larvae fed enriched Artemia . Survival rate, intermolt period, carapace width at the first crab stage, ratio of chela to carapace length at the fifth zoeal stage and the occurrence of molting failure to megalops were observed. Mud crab larvae were reared in 1-L plastic beakers and fed with Artemia enriched at five different levels of EPA (0.31% to 1.36% EPA; referred to as E25, E50, E75 and E100) or four different levels of DHA (0.75–0.95% EPA and 0.49–1.38% DHA; referred to as D25, D50, D75 and D100). As a negative control, larvae were fed Artemia enriched with oleic acid (OA). Mud crab larvae fed Artemia containing low (0.41–0.45% EPA and trace DHA) or high (1.36% and 0.95% EPA and 0.16% and 1.38% DHA) amounts of EPA and DHA showed a significantly lower survival rate and prolonged intermolt period ( P  < 0.05). Moreover, a high frequency of molting failure to the megalops stage (34 and 33%) occurs coincident with a high chela to carapace length ratio (43 and 44%) in mud crab larvae fed high amounts of EPA and DHA (E100 and D100), suggesting that both of these treatments contain EPA and DHA in excess. These results indicate that during Artemia feeding, EPA and DHA content should be adjusted to 0.71–0.87% and 0.49–0.72% for maintaining a high survival, accelerating the intermolt period, and producing larger carapace width in the first crab stage.     

Formation and Deposition of Ozone in a Red Pine Forest

Concentrations of ozone and nitrogen oxides, together with air temperature and solar radiation intensity, were measured at several heights on a tower standing through the canopy of a red pine forest in summer and in autumn. In the summer observation, the diurnal variation patterns of ozone concentration both above and below the canopy were all similar and parallel to the solar radiation intensity. Using the data collected immediately above the canopy, deviation from the Leighton relationship and variations of concentration sums [O₃] + [NO] and [NO₂] + [NO] were examined, and as a result, it was supposedthat ozone was photochemically formed there in the daytime, probably because hydrocarbons emitted from pine trees broke the photostationary state among ozone and nitrogen oxides. The vertical temperature profile exhibited an inversion at the leaf-layer, which must have hindered vertical mixing of the air and made the trunk space more or less isolated from the upper atmosphere. These observations led to an idea that the similarity of the ozone variation pattern at every height was caused by the photochemical formation that proceeded simultaneously above and below the canopy rather than by vertical transport. Such situations of ozone formation were supported by observation of two maximums in the ozone vertical profile, one immediately above the canopy and another in the trunk space. Another feature of the ozone profile was a deep minimum in the leaf layer, which indicated ozone deposition onto leaf surfaces. This study thus revealed concurrence of ozone formation and deposition, and left two potentially important implications worthy of further investigation: (1) a forest is not always a sink but can be a source of ozone in sunlit conditions, and (2) deposition of ozone to trees can take place not only from outside but also from inside of a forest. In the autumn observation, however, the ozone formation was barely recognizable above the canopy and no longer found in the trunk space; in addition, the ozone concentration minimum in the leaf layer disappeared, suggesting that the deposition or removal was dependent on temperature.     

Diffuse alveolar damage in a young cat

A 10-month-old cat was diagnosed with congenital subvalvular aortic stenosis. To resolve its hypoxia, oxygen therapy was administered a couple of times a week during two months. The oxygen partial pressure in the chamber was maintained between 30 and 35%, and the time for one procedure was 12-24 hr. The animal died due to severe respiratory failure. At necropsy, the lungs were voluminous and had a rubbery texture. Histologically, large type II pneumocytes with occasional atypia had diffusely proliferated within the lungs. Interstitial fibrosis was not observed, although some alveolar septa were thickened along with fibrinous exudates and neutrophilic infiltration. The histology of these lesions was consistent with diffuse alveolar damage (DAD), which might have been partially due to oxygen toxicity.