Regulation of palmitoyl-CoA chain elongation and linoleoyl-CoA chain elongation in rat liver microsomes and the differential effects of peroxisome proliferators, insulin and thyroid hormone

Treatment of rats with p-chlorophenoxyisobutyric acid (clofibric acid), 2,2'-(decamethylenedithio)diethanol, di(2-ethylhexyl)phthalate or acetylsalicylic acid caused an increase in activity of palmitoyl-CoA chain elongation in hepatic microsomes. The activity of palmitoyl-CoA chain elongation decreased in both hypothyroid-state and diabetic-state rats, increased in hyperthyroid-state rats and did not change in adrenalectomized rats. The administration of clofibric acid to these rats in an altered hormonal state caused an increase in the activity of palmitoyl-CoA chain elongation, but no additional increase in the activity was observed with treatment of hyperthyroid rats with clofibric acid. The activity of linoleoyl-CoA chain elongation did not respond to the changes in either the nutritional conditions or the hormonal state of insulin so sensitively as the activity of palmitoyl-CoA chain elongation. The treatment of rats with triiodothyronine caused a marked increase in the activity of linoleoyl-CoA chain elongation; nevertheless, the activity of linoleoyl-CoA chain elongation was not changed by the treatment of rats with clofibric acid. The results suggest that rat liver microsomes contain at least two fatty acid chain elongation systems and that these chain elongation systems are regulated differently by hormones and drugs.     

Genetic control of petiole length in Arabidopsis thaliana

Shade-avoidance syndrome is characterized by the formation of elongated petioles and unexpanded leaf blades under low-intensity light, but the genetic basis for these responses is unknown. In this study, two-dimensional mutational analysis revealed that the gene for phytochrome B, PHYB, had opposing effects in the leaf petioles and leaf blades of Arabidopsis, while the ROT3, ACL2, and GAI genes influenced the length of leaf petioles more significantly than the length of leaf blades. Anatomical analysis revealed that the PHYB and ACL2 genes control the length of leaf petioles exclusively via control of the length of individual cells, while the GAI, GA1 and ROT3 genes appeared to control both the elongation and proliferation of petiole cells, in particular, under strong light. By contrast, both the size and the number of cells were affected by the mutations examined in leaf blades. The differential control of leaf petiole length and leaf blade expansion is discussed.     

Macrophage Glucose-6-Phosphate Dehydrogenase Stimulates Proinflammatory Responses with Oxidative Stress

Glucose-6-phosphate dehydrogenase (G6PD) is a key enzyme that regulates cellular redox potential. In this study, we demonstrate that macrophage G6PD plays an important role in the modulation of proinflammatory responses and oxidative stress. The G6PD levels in macrophages in the adipose tissue of obese animals were elevated, and G6PD mRNA levels positively correlated with those of proinflammatory genes. Lipopolysaccharide (LPS) and free fatty acids, which initiate proinflammatory signals, stimulated macrophage G6PD. Overexpression of macrophage G6PD potentiated the expression of proinflammatory and pro-oxidative genes responsible for the aggravation of insulin sensitivity in adipocytes. In contrast, when macrophage G6PD was inhibited or suppressed via chemical inhibitors or small interfering RNA (siRNA), respectively, basal and LPS-induced proinflammatory gene expression was attenuated. Furthermore, macrophage G6PD increased activation of the p38 mitogen-activated protein kinase (MAPK) and NF-κB pathways, which may lead to a vicious cycle of oxidative stress and proinflammatory cascade. Together, these data suggest that an abnormal increase of G6PD in macrophages promotes oxidative stress and inflammatory responses in the adipose tissue of obese animals.     

Changes in brain energy metabolism,neurotransmitters, and choline during and after incomplete cerebral ischemia in spontaneously hypertensive rats

In order to investigate changes in energy metabolism, neurotransmitters, and membrane disorder accompanying incomplete cerebral ischemia, a bilateral common carotid artery occlusion model of spontaneously hypertensive rats was utilized. We measured concentrations of ATP, phosphocreatine (PCr), lactate (Lac), glucose (Glu), acetylcholine (ACh), choline (Ch), and -aminobutyric acid (GABA) in both the cerebral cortex and the subcortical regions after 1 h ischemia, 2 h ischemia, and 2 h reflow following 2 h ischemia, and then examined changes in concentrations of these substances during and after incomplete cerebral ischemia. Also examined were interrelations of changes in these substance levels during ischemia. In the cerebral cortex, levels of ATP, PCr, Glu, and ACh decreased, and levels of Lac, Ch, and GABA increased during ischemia. After recirculation, levels of ATP, PCr, Ch, and GABA tended to return to the normal range. On the other hand, the Lac level remained in the ischemic range and the Glu level rose and greatly exceeded the normal range. With regard to ACh, most animals showed normal levels but some exceeded the normal range. Changes in the subcortical regions were qualitatively the same as those in the cerebral cortex during and after ischemia (except with Glu), but only smaller in degrees. Glu levels remained unchanged during ischemia. Correlation of the levels of these substances in the cerebral cortex was examined using normal and ischemic values. A high correlation was generally observed between ATP and other substance levels. The relations between ATP and either PCr or Glu levels were linear. The relation between ATP and ACh levels was logarithmic. The relations between ATP and either Lac, Ch, or GABA levels were exponential. Namely, ACh, Lac, Ch, and GABA levels stayed constant until ATP fell to some fixed low level, suggesting the existence of a threshold. High correlations were also observed among Lac, Ch, and GABA levels.     

The Arabidopsis cucumovirus multiplication 1 and 2 loci encode translation initiation factors 4E and 4G

The cum1 and cum2 mutations of Arabidopsis thaliana inhibit cucumber mosaic virus (CMV) multiplication. In cum1 and cum2 protoplasts, CMV RNA and the coat protein accumulated to wild-type levels, but the accumulation of the 3a protein of CMV, which is necessary for cell-to-cell movement of the virus, was strongly reduced compared with that in wild-type protoplasts. In cum2 protoplasts, the accumulation of turnip crinkle virus (TCV)-related RNA and proteins was also reduced. Positional cloning demonstrated that CUM1 and CUM2 encode eukaryotic translation initiation factors 4E and 4G, respectively. Unlike most cellular mRNA, the CMV RNA lacks a poly(A) tail, whereas the TCV RNA lacks both a 5′-terminal cap and a poly(A) tail. In vivo translation analyses, using chimeric luciferase mRNA carrying the terminal structures and untranslated sequences of the CMV or TCV RNA, demonstrated that these viral untranslated sequences contain elements that regulate the expression of encoded proteins positively or negatively. The cum1 and cum2 mutations had different effects on the action of these elements, suggesting that the cum1 and cum2 mutations cause inefficient production of CMV 3a protein and that the cum2 mutation affects the production of TCV-encoded proteins.     

Tissue-autonomous promotion of palisade cell development by phototropin 2 in Arabidopsis

Light is an important environmental information source that plants use to modify their growth and development. Palisade parenchyma cells in leaves develop cylindrical shapes in response to blue light; however, the photosensory mechanism for this response has not been elucidated. In this study, we analyzed the palisade cell response in phototropin-deficient mutants. First, we found that two different light-sensing mechanisms contributed to the response in different proportions depending on the light intensity. One response observed under lower intensities of blue light was mediated exclusively by a blue light photoreceptor, phototropin 2 (PHOT2). Another response was elicited under higher intensities of light in a phototropin-independent manner. To determine the tissue in which PHOT2 perceives the light stimulus to regulate the response, green fluorescent protein (GFP)-tagged PHOT2 (P2G) was expressed under the control of tissue-specific promoters in the phot1 phot2 mutant background. The results revealed that the expression of P2G in the mesophyll, but not in the epidermis, promoted palisade cell development. Furthermore, a constitutively active C-terminal kinase fragment of PHOT2 fused to GFP (P2CG) promoted the development of cylindrical palisade cells in the proper direction without the directional cue provided by light. Hence, in response to blue light, PHOT2 promotes the development of cylindrical palisade cells along a predetermined axis in a tissue-autonomous manner.     

The targeted disruption of the CD98 gene results in embryonic lethality

CD98 is one of the important molecules for development, cell differentiation, cell proliferation, and regulation of cellular function. In this study, CD98 heavy chain (HC) knockout mice were produced and analyzed. Five targeted ES clones were obtained and colony frequency was about 2%. One (clone 113) of the five heterozygous ES cell clones had undergone aberrant recombination at the 5' side. The aberrant recombination happened at the site between second intron and 5' arm. All lines from correctly targeted clones could not transmit the mutated allele to spermatozoa. The mutated allele derived from the aberrant targeted clone was transmitted to the progeny. However, none of the F2 mice was homozygous for the CD98 mutation, indicating that the targeted disruption of the CD98 gene results in embryonic lethality. The point of embryonic lethality is considered to be between 3.5 and 9.5 dps. These findings indicate that CD98 molecules are essential for mouse embryogenesis.     

Decreased particulate NADH oxidase activity in Bacillus subtilis spores after polymyxin B treatment

The activities of several enzymes of polymyxin B-treated dormant and germinated spores of Bacillus subtilis were examined. The particulate NADH oxidase of the antibiotic-treated spores showed considerably lower specific and total activities compared with those of untreated ones. The specific and total NADH oxidase activities of untreated spores increased 12- and 15-fold respectively during germination, whereas increases during germination of polymyxin B-treated spores were inhibited. The specific and total activities of particulate NADH cytochrome c reductase of dormant spores were decreased by polymyxin B treatment in almost the same proportion as those of the particulate NADH oxidase. The specific activity of NADH dehydrogenase of dormant spores remained unchanged after antibiotic treatment but the total activity fell considerably. The activities of other enzymes examined were similar for untreated dormant and germinated spores and antibiotic-treated spores. The respiration of polymyxin B-treated dormant spores was inhibited at the same time as the start of germination. Morphologically, polymyxin B-treated dormant spores lost a laminar structure of the cortex and details of the spore protoplast. The inhibitory mechanism of particulate NADH oxidase activity of polymyxin B-treated dormant spores is discussed.     

Sex-related difference in the inductions by perfluoro-octanoic acid of peroxisomal beta-oxidation, microsomal 1-acylglycerophosphocholine acyltransferase and cytosolic long-chain acyl-CoA hydrolase in rat liver.

Inductions by perfluoro-octanoic acid (PFOA) of hepatomegaly, peroxisomal beta-oxidation, microsomal 1-acylglycerophosphocholine acyltransferase and cytosolic long-chain acyl-CoA hydrolase were compared in liver between male and female rats. Marked inductions of these four parameters were seen concurrently in liver of male rats, whereas the inductions in liver of female rats were far less pronounced. The sex-related difference in the response of rat liver to PFOA was much more marked than that seen with p-chlorophenoxyisobutyric acid (clofibric acid) or 2,2'-(decamethylenedithio)diethanol (tiadenol). Hormonal manipulations revealed that this sex-related difference in the inductions is strongly dependent on sex hormones, namely that testosterone is necessary for the inductions, whereas oestradiol prevented the inductions by PFOA.