Chromosome banding in the genusPinus

Somatic chromosomes (2n=24) ofPinus luchuensis Mayr at metaphase were observed by fluorescent banding methods with chromomycin A₃ (CMA) and DAPI. CMA-bands appeared at the interstitial and/or proximal regions of nearly all chromosomes. DAPI-bands appeared at the interstitial and/or centromeric regions of nearly all chromosomes, and pairs of DAPI-dots appeared at the centromeric regions. Each homologous pair of chromosomes in the chromosome complement was identified by the CMA and DAPI fluorescent banding patterns. The interstitial CMA-bands were mostly localized at the secondary constrictions of the Feulgen-stained chromosomes. The fluorescent banding pattern ofP. luchuensis was very similar to that ofP. thunbergii, but was different from that ofP. densiflora.     

A novel protein kinase B (PKB)/AKT-binding protein enhances PKB kinase activity and regulates DNA synthesis

Protein kinase B (PKB)/Akt reportedly plays a role in the survival and/or proliferation of cells. We identified a novel protein, which binds to PKB, using a yeast two-hybrid screening system. This association was demonstrated not only in vivo by overexpressing both proteins or by coimmunoprecipitation of the endogenous proteins, but also in vitro using glutathione S-transferase fusion proteins. Importantly, this protein specifically associates with the C terminus of PKB but not with other AGC kinases and enhances PKB phosphorylation and kinase activation without growth factor stimulation. Thus, we termed this Akt-specific binding protein APE (Akt-phosphorylation enhancer). Since APE-induced phosphorylation of PKB did not occur in cells treated with wortmannin or LY294002, APE itself is not a kinase but seems to enhance or prolong the phosphoinositide 3-kinase-dependent phosphorylation of PKB. In cells in which APE was suppressed by small interfering RNA, DNA synthesis was significantly reduced with suppression of PKB phosphorylation, suggesting a synergistic role of APE in PKB-induced proliferation. On the other hand, in cells overexpressing both PKB and APE, despite markedly increased basal phosphorylation of PKB, both DNA rereplication and subsequent Chk2 phosphorylation and apoptosis were seen, suggesting the involvement of APE in the regulation of cell cycling replication licensing. Taking these observations together, APE appears to be a novel regulator of PKB phosphorylation. Furthermore, the interaction between APE and PKB, possibly dependent on the expression levels of both proteins, may be a novel molecular mechanism leading to proliferation and/or apoptosis.     

Three mitogen-activated protein kinases inhibit insulin signaling by different mechanisms in 3T3-L1 adipocytes

TNFalpha, which activates three different MAPKs [ERK, p38, and jun amino terminal kinase (JNK)], also induces insulin resistance. To better understand the respective roles of these three MAPK pathways in insulin signaling and their contribution to insulin resistance, constitutively active MAPK/ERK kinase (MEK)1, MAPK kinase (MKK6), and MKK7 mutants were overexpressed in 3T3-L1 adipocytes using an adenovirus-mediated transfection procedure. The MEK1 mutant, which activates ERK, markedly down-regulated expression of the insulin receptor (IR) and its major substrates, IRS-1 and IRS-2, mRNA and protein, and in turn reduced tyrosine phosphorylation of IR as well as IRS-1 and IRS-2 and their associated phosphatidyl inositol 3-kinase (PI3K) activity. The MKK6 mutant, which activates p38, moderately inhibited IRS-1 and IRS-2 expressions and IRS-1-associated PI3K activity without exerting a significant effect on the IR. Finally, the MKK7 mutant, which activates JNK, reduced tyrosine phosphorylation of IRS-1 and IRS-2 and IRS-associated PI3K activity without affecting expression of the IR, IRS-1, or IRS-2. In the context of our earlier report showing down-regulation of glucose transporter 4 by MEK1-ERK and MKK6/3-p38, the present findings suggest that chronic activation of ERK, p38, or JNK can induce insulin resistance by affecting glucose transporter expression and insulin signaling, though via distinctly different mechanisms. The contribution of ERK is, however, the strongest.     

Hydrothermal conversion of municipal organic waste into resources

Sub- and supercritical water have been focused on as an environmentally attractive reaction media where organic materials can be decomposed into smaller molecules. We applied a hydrothermal reaction in subcritical water to the treatment of rabbit food as a model municipal solid waste. The reaction was carried out in a batch reactor at the temperature range of 473-623 K or in a semi-continuous reactor with the temperature profile from 473 to 573 K. The liquid reaction products were separated into water-soluble and water-insoluble parts. The water-soluble part was analyzed in terms of glucose and organic acids. For the batch reactor, the largest amount of water-soluble component was approximately 50%. Major organic acids detected were acetic acid and lactic acid. The largest yields were 2.6% for acetic acid and 3.2% for lactic acid. The largest yield of glucose was 33% at 523 K. For the semi-continuous reactor, the glucose yields were 11.5%, 3.9% and 8.7% in each fraction obtained at 473, 523, and 573 K, respectively.     

Pressurized hot water extraction of anthraquinones from the roots of Morinda citrifolia

This study examines pressurized hot water extraction of anthraquinones from dried roots of Morinda citrifolia (Noni). The effects of various operating conditions such as water temperature (110, 170, and 220 degrees C), and water flow rate (2, 4, and 6 mL min(-1)) on extraction yield and extraction rate were determined. At 220 degrees C, the extraction yield was the highest and was approximately 43.6 mg g(-1). Subcritical water extraction at 4 mL min(-1) or higher was found to be able to recover all the antraquinones present in the roots within 3 h of extraction, even at 170 degrees C. Pressure had no significant effect on the results for the range of temperatures studied. The flow rate of 6 mL min(-1) resulted in the highest extraction rate, but the extraction efficiency, as measured by the amount of anthraquinones extracted per unit volume of water was lower than that of 4 mL min(-1). Anthraquinones solubility in pressurized hot water was determined. The results indicate a presence of a mass transfer limitation in the extraction process from solid matrix.     

Recognition of tennis serve performed by a digital player: comparison among polygon, shadow, and stick-figure models

The objective of this study was to assess the cognitive effect of human character models on the observer's ability to extract relevant information from computer graphics animation of tennis serve motions. Three digital human models (polygon, shadow, and stick-figure) were used to display the computationally simulated serve motions, which were perturbed at the racket-arm by modulating the speed (slower or faster) of one of the joint rotations (wrist, elbow, or shoulder). Twenty-one experienced tennis players and 21 novices made discrimination responses about the modulated joint and also specified the perceived swing speeds on a visual analogue scale. The result showed that the discrimination accuracies of the experienced players were both above and below chance level depending on the modulated joint whereas those of the novices mostly remained at chance or guessing levels. As far as the experienced players were concerned, the polygon model decreased the discrimination accuracy as compared with the stick-figure model. This suggests that the complicated pictorial information may have a distracting effect on the recognition of the observed action. On the other hand, the perceived swing speed of the perturbed motion relative to the control was lower for the stick-figure model than for the polygon model regardless of the skill level. This result suggests that the simplified visual information can bias the perception of the motion speed toward slower. It was also shown that the increasing the joint rotation speed increased the perceived swing speed, although the resulting racket velocity had little correlation with this speed sensation. Collectively, observer's recognition of the motion pattern and perception of the motion speed can be affected by the pictorial information of the human model as well as by the perturbation processing applied to the observed motion.     

DNA ploidy and proliferative activity of human pulmonary epithelium

DNA ploidy and distribution has been determined in normal and abnormal bronchial, bronchiolar and alveolar epithelium from 22 patients, aged between 0 and 85 years, 9 of whom had received chemotherapy for malignant disease. The DNA ploidy was diploid in all the specimens examined. The S + G2/M fraction was significantly greater in diseased than normal bronchial trees. In the bronchial epithelium, mean values +/- the standard deviation (SD) were 5.5 +/- 2.2% vs 1.1 +/- 0.6%, in bronchiolar epithelium 4.6 +/- 1.6% vs 1.0 +/- 0.9% and in alveolar epithelium 4.6 +/- 1.6% vs 0.8 +/- 0.5%. The highest S + G2/M value of 8.9% was obtained from inflamed bronchial epithelium. Polyploid cells up to the octaploid range occurred infrequently but their incidence was slightly increased to between 0.16% and 0.9% in diseased lungs and in patients who had received chemotherapeutic drugs. It was concluded that (1) non-cancerous drugs. It was concluded that (1) non-cancerous pulmonary epithelium is diploid, that (2) pulmonary epithelium shows steady-state renewal at all ages and polyploid cells are rare under normal conditions and that (3) the S + G2/M fraction increases up to approximately 10% in reactive proliferative states.     

Physicochemical measurements of Japonica rice cultivars in Heilongjiang Province

We here characterized 27 japonica rice cultivars grown in Heilongjiang province and evaluated the relationship among their iodine absorption curve, physical properties, and ratio of 13 kDa prolamin. We developed the novel estimation formulae for ratio of 13 kDa prolamin and overall hardness (H2) with the use of A_λmax and λmax.     

Thermoperiodism Mechanism in the Germination of Cocklebur Seeds

In thermoperiodic germination of secondarily dormant cocklebur(Xanthium pennsylvanicum Wallr.) seeds, the extent of electronflow through the CN-sensitive, cytochrome path and the CN-resistant,alternative path differred in the cool and warm phases. Thecool phase triggered active engagement of the alternative pathduring the subsequent warm phase, and this led to an increasein the ratio of the alternative path flux to the cytochromepath flux. The cool and warm phases had different functionsin adenylate metabolism. The former acted mainly in the productionand accumulation of ATP, whereas the latter supplied ADP and,especially, AMP. Thus an increasing number of sequential thermoperiodiccycles augmented the size of the adenylate pool and the energycharge, both of which may be necessary for germination to takeplace. (Received September 16, 1981; Accepted November 5, 1982)     

Nocturnal enhancement of plasma melatonin could be suppressed by benzodiazepines in humans

Plasma melatonin levels were determined every 20 and 30 min for 24 hours on the last day of repeated oral administrations (1 or 2 mg a day for 8 or 9 days) of a benzodiazepine derivative (450191-s), which is known to be metabolized to active benzodiazepines after administration. In one of the two subjects, the nocturnal enhancement of plasma melatonin which was obvious on a control day with placebo was diminished almost completely. In the other subject, observed were not only the diminishment of its nocturnal enhancement but also its increase during the daytime almost to the nocturnal levels on a control day, which may indicate a rebound increase in melatonin synthesis or a shift in its day-night rhythmicity. Such suppressing effects of benzodiazepines on the nocturnal plasma melatonin levels were also examined in the case of a single administration of 2 mg of 450191-s or flunitrazepam in the second series of experiments. Even a single flunitrazepam seemed to have lowered nocturnal plasma melatonin levels, which then recovered to the usual levels following the administration of 5 mg of a benzodiazepine antagonist, Ro 15-1788, given 6 hours after the flunitrazepam. However, single 450191-s did not show any remarkable effects. Thus, it has been suggested that benzodiazepines could suppress the nocturnal levels of plasma melatonin or shift its day-night rhythmicity at least when administered repeatedly. The possible action site of benzodiazepines may be the central nervous system, since melatonin synthesis has been though to be under strongly regulated by the central nervous pathway from the retina to the pineal body. Therefore, these effects of benzodiazepines may provide a method for investigating the physiological role of melatonin and its day-night rhythmicity as well as to further clarify the system regulating melatonin synthesis in humans.