Neonatal presentation of adult-onset type II citrullinemia

Adult-onset type II citrullinemia (CTLN2) is characterized by a liver-specific argininosuccinate synthetase deficiency caused by a deficiency of the citrin protein encoded by the SLC25A13 gene. Until now, however, no SLC25A13 mutations have been reported in children with liver diseases. We described three infants who presented as neonates with intrahepatic cholestasis associated with hypermethioninemia or hypergalactosemia detected by neonatal mass screening. DNA analyses of SLC25A13 revealed that one patient was a compound heterozygote for the 851de14 and IVS11+IG-->A mutations and two patients (siblings) were homozygotes for the IVS11+lG-->A mutation. These results suggested that there may be a variety of liver diseases related to CTLN2 in children.     

Silicon intake to vertebral columns of mice after dietary supply

Vertebral columns were dissected and analyzed after birth with oral administration of silicon for 4 wk and for 8 wk. The silicon level was lower (20 μg/g) at the beginning. It remains unchanged after 4 wk and then increases twice as much as that for those mice bred for 8 wk than those bred for 4 wk. This increase depends remarkably on the mass ratio of Si/Ca (M/M). The ratio increases to three times higher than that of the control at the beginning of the experiments (5 wk after birth). Although the S and P contents appeared to be lower, these increased when Si was administered in combination with phosphopeptide. Other elements, such as Ca, Mg, Fe, and Zn, appeared to be unchanged as the weeks proceeded. These findings seem to support a proposal that silicon is necessary for the growth of backbones in mice.     

草酰乙酸和苹果酸对菠菜叶片和完整叶绿体光合作用的影响

观测了OAA和MA对菠菜叶片和完整叶绿体光合作用的影响.结果显示,当叶片切块在20μmol/L的OAA存在时,其叶片的光合放氧速率增加了89%,经OAA处理的离体完整叶绿体的光合放氧速率增加了72%;当反应体系中存在有较高浓度的NaHCO3时,OAA的作用不明显.叶片经20 μmol/L的MA处理后,叶片光合放氧速率比对照高127%.用CO2分析仪观测了处理后叶片的净光合速率(Pn),结果显示,OAA和MA处理后的叶片Pn值分别是对照的117%和111%.对在C3植物中建立C4微循环系统来提高光合作用效率的可能性进行了讨论.     

The complete nucleotide sequence of the mitochondrial genome of Tetraodon nigroviridis.

The fresh water pufferfish Tetraodon nigroviridis is a model organism for studying evolution of genome and gene functions, but its mitochondrial genome (mtDNA) sequence is still not available. We determined the complete nucleotide sequence of its mtDNA using shotgun sequencing. The T. nigroviridis mtDNA was 16,462 bp, and contained 13 protein coding genes, 22 tRNAs, 2 rRNAs and a major non-coding region. The gene order was identical to the common type of vertebrate mtDNA, whereas the G + C content in the sense strand was 46.9%, much higher than most other fish species. One hundred and three SNPs were detected in the control region of the mtDNA of 35 individuals, a majority of SNPs were detected in the 5' end of the control region. A phylogenetic study including 21 fish species was performed on concatenated amino acid sequences of 12 protein coding genes, and revealed that the T. nigroviridis was clustered with Fugu rubripes into a group. The complete mtDNA sequence and SNPs in its control region will be useful in studying fish evolution, in differentiating different Tetraodon species and in analyzing genetic diversity within T. nigroviridis.     

Inhibition of brain prostaglandin D synthetase and prostaglandin D2 dehydrogenase by some saturated and unsaturated fatty acids

The activities of rat brain prostaglandin D synthetase and swine brain prostaglandin D2 dehydrogenase were inhibited by some saturated and unsaturated fatty acids. Myristic acid was most potent among saturated straight-chain fatty acids so far tested. The IC50 values of this acid were 80 microM for prostaglandin D synthetase and 7 microM for prostaglandin D2 dehydrogenase, respectively. Little inhibition was found with methyl myristate and myristyl alcohol. The IC50 values of these derivatives were more than 200 microM for both enzymes, suggesting that the free carboxyl group was essential for the inhibition. The effects of cis double bond structure of fatty acids on the inhibition potency were examined by the use of the carbon 18 and 20 fatty acids. The inhibition potencies for both enzymes increased with the number of cis double bonds; the IC50 values of stearic, oleic, linoleic and linolenic acid were, respectively, more than 200, 60, 30 and 30 microM for prostaglandin D synthetase, and 20, 10, 8.5 and 7 microM for prostaglandin D2 dehydrogenase. Arachidonic acid also inhibited the activities of both enzymes with respective IC50 values of 40 microM for prostaglandin D synthetase and 3.9 microM for prostaglandin D2 dehydrogenase, while arachidic acid showed little inhibition. The kinetic studies with myristic acid and arachidonic acid demonstrated that the inhibition by these fatty acids was competitive and reversible for both enzymes. Myristic acid and other fatty acids also inhibited the activities of several enzymes in prostaglandin metabolism, although to a lesser extent. The IC50 values of myristic acid for prostaglandin E isomerase, thromboxane synthetase and NAD-linked prostaglandin dehydrogenase (type I) were 200, 700 and 100 microM, respectively. However, this fatty acid showed little inhibition on fatty acid cyclooxygenase (20% at 800 microM), glutathione-requiring prostaglandin D synthetase from rat spleen (20% at 800 microM), and NADP-linked prostaglandin dehydrogenase (type II) (no inhibition at 200 microM).     

Molecular characterization,relatedness and antifungal susceptibility of the basidiomycetous hormographiella species and Coprinus cinereus from clinical and environmental sources

Hormographiella-like strains, isolated from different natural substrates and producing sclerotia and occasionally basidiomata of Coprinus cinereus, were compared morphologically and using molecular techniques with clinical strains of Hormographiella aspergillata and H. verticillata. Analysis of restriction fragment length polymorphisms of ribosomal and mitochondrial-like DNA confirmed interspecific differences between H. aspergillata and H. verticillata, supporting the morphological data, and helped demonstrate that H. aspergillata is the anamorph of C. cinereus. The latter was confirmed also by crossing tests. The analysis of the mtDNA restriction profiles revealed intraspecific variability in C. cinereus, which allowed differentiation of clinical and environmental strains. Due to the implication of C. cinereus and Hormographiella in human opportunistic infections, the antifungal susceptibility test is included. Results show that all strains were susceptible to miconazole, itraconazole and ketoconazole but not to flucytosine and fluconazol. Susceptibility against amphotericin B was variable; while H. verticillata was susceptible, four out of seven C. cinereus strains tested were resistant.     

Increased expression in vivo and in vitro of foreign genes directed by A-type inclusion body hybrid promoters in recombinant vaccinia viruses.

We constructed A-type inclusion body (ATI) hybrid promoters, that is, late ATI promoters followed by tandemly repeated early regions of the promoter for the 7.5-kDa protein (the 7.5-kDa promoter). The repetition of the whole early promoter sequence of the 7.5-kDa gene, including the upstream consensus sequence and initiation region, efficiently increased the early expression of the bacterial chloramphenicol acetyltransferase gene in recombinant vaccinia virus. Recombinant vaccinia virus could express influenza virus hemagglutinin via the hybrid promoter more efficiently, induced higher levels of neutralizing antibody and cytotoxic T lymphocytes, and consequently protected mice more efficiently against challenge with influenza virus than did recombinant vaccinia virus containing the widely used 7.5-kDa promoter.     

Cochliomycin A inhibits the larval settlement of Amphibalanus amphitrite by activating the NO/cGMP pathway

Cochliomycin A is a compound with anti-barnacle settlement activity and low toxicity, but the molecular mechanism of the compound is unknown. Here, isobaric tags for the relative or absolute quantitation (iTRAQ) labeling proteomic method were applied to analyze changes in the proteome of Amphibalanus（=Balanus） amphitrite cyprids in response to cochliomycin A treatment. Cochliomycin A affected the cytochrome P450, glutathione S-transferase (GST) and NO/cGMP pathways, among which the NO/cGMP pathway was considered to play a key role in barnacle larval settlement, while the cytochrome P450 and the GST pathways are mainly for detoxification. The results of real-time PCR further suggested the NO/cGMP pathway was activated in response to cochliomycin A. Larval settlement assays revealed that S-methylisothiourea sulfate (SMIS) and 1H-(1,2,4)oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) rescued cyprids from cochliomycin A-induced inhibition of larval settlement. The findings supported the hypothesis that cochliomycin A inhibited barnacle larval settlement by stimulating the NO/cGMP pathway.     

Low-temperature dynamic light scattering. I. Structural reorganization and physical gel formation in cellulose triacetate/methyl acetate dilute solution at -99 - 45 degrees C

Curious low-temperature solubility of cellulose triacetates (CTA; here we use nominally "CTA," but the sample still contains 7% of C-6 position hydroxyls) in an organic solvent, methyl acetate (MA), was studied by a newly designed low-temperature type of DLS apparatus, which enabled for the first time to investigate the structural change of CTA in solution from 45 degrees C down to -100 degrees C. A molecularly dissolved CTA was found to coexist with three types of self-assemblies over all the temperature ranges except for the three specific temperatures T* of 30, -10, and -75 degrees C. However, these multiple self-assemblies are not in real thermodynamic equilibrium but in a metastable state, which could be stabilized effectively by the intermolecular hydrogen bonding (HB) with the help of the dipole interaction at low temperatures. In more detail, with decreasing temperature, these assemblies performed the structural reorganization drastically at three T*'s and would finally be frozen in a physical gel structure at -99 degrees C; around the freezing temperature of MA, CTA molecules could be trapped homogeneously in the frozen MA. The crucial role in such structural reorganizations is played by the balance between the intermolecular HB and the dipole interaction worked in the highly electronegative solvent. Because these interactions, which are mediated by the solvent electronegativity, change drastically with temperature, they result in the control of not only the single CTA chain conformation (= the intramolecular HB) but also the binding ways of the intermolecular HBs between CTA molecules and they induce multitudinous metastable structures in solution. Here it is noted that HB could work mainly between the C-6 position hydroxyls in the anhydroglucose units of CTA and are essentially effective at low temperatures.