Pretreatment and posttreatment in the biolistic transformation of tea plant (Camellia sinensis) somatic embryos.

The tea plant (Camellia sinensis) contains various metabolic substances, including catechins and caffeine, for which genetic transformation techniques are essential for investigating the associated metabolic pathways. In this study, we sought to optimize the conditions and culture process for particle bombardment-mediated transformation of tea plant somatic embryos. We describe somatic embryo pretreatment for effective transient transformation in biolistic bombardment and the posttreatment conditions of somatic embryos for accelerating differentiation after bombardment. For the purpose of transformation, we used the somatic embryos of C. sinensis var. assamica 'Tingamira normal,' which were cultured in Murashige and Skoog (MS) medium containing 2 mg l-1 indole-3-butyric acid (IBA) and 4 mg l-1 6-benzyladenine (BA) at 25°C ±2°C under a 16-h photoperiod. With respect to the optimization of particle bombardment conditions for tea somatic embryos, we examined the effects of different Au colloid particle diameters and bombardment pressures, and accordingly established bombardment with 1.0-µm-diameter DNA-coated Au colloid at 1,100 psi as optimal conditions for introducing DNA for the transient expression of GUS. Additionally, we found that transplantation of tea somatic embryos from IBA/BA medium to a hormone-free medium prior to bombardment and incubation in the dark post-bombardment increased the frequency of secondary embryo production. Furthermore, osmotic treatment by culturing the somatic embryos in medium supplemented with 0.4 M mannitol improved transient transformation efficiency. After transformation, the culture of somatic embryos on filter papers or Kimwipes soaked in MS medium facilitated rapid and effective development of the somatic embryos.

A magnetic carbon sorbent for radioactive material from the Fukushima nuclear accident.

Here we present the first report of a carbon-γ-Fe2O3 nanoparticle composite of mesoporous carbon, bearing COOH- and phenolic OH- functional groups on its surface, a remarkable and magnetically separable adsorbent, for the radioactive material emitted by the Fukushima Daiichi nuclear power plant accident. Contaminated water and soil at a level of 1,739 Bq kg(-1) ((134)Cs and (137)Cs at 509 Bq kg(-1) and 1,230 Bq kg(-1), respectively) and 114,000 Bq kg(-1) ((134)Cs and (137)Cs at 38,700 Bq kg(-1) and 75,300 Bq kg(-1), respectively) were decontaminated by 99% and 90% respectively with just one treatment carried out in Nihonmatsu city in Fukushima. Since this material is remarkably high performance, magnetically separable, and a readily applicable technology, it would reduce the environmental impact of the Fukushima accident if it were used.

Usefulness of fibrin degradation products and d-dimer levels as biomarkers that reflect the severity of trauma.

PURPOSE: To evaluate the usefulness of the levels of fibrin degradation products (FDPs) and d-dimer (DD) in blood for reflecting the severity of trauma. PATIENTS AND METHODS: We reviewed medical records of trauma patients who were transferred to the emergency department of Gunma University Hospital between January 2010 and December 2010. The relationships among Injury Severity Score, mean blood pressure, heart rate, hemoglobin, hematocrit, platelet count, fibrinogen, international normalized ratio of prothrombin time, activated partial thromboplastin time, FDP, and DD on arrival were examined in those patients. RESULTS: A total of 122 patients were included in this study. The coefficients of correlation of FDP (r = 0.710) and DD (r = 0.636) with Injury Severity Score were higher than those of other parameters. In addition, the areas under the receiver operating characteristic curve of FDP and DD were larger than those of other parameters (0.757 and 0.756, respectively). The cutoff value of FDP from the Youden index was 4.70 µg/mL, and the sensitivity and specificity values were 75.9% and 68.4%, respectively. The cutoff value of DD from the Youden index was 2.55 µg/mL, and the sensitivity and specificity values were 75.9% and 73.7%, respectively. There were four patients requiring more than 10 U of red blood cell transfusion within 24 hours after trauma (a unit of red blood cell transfusion is made from 200 mL of whole blood in Japan). In those four patients, the mean values of FDP and DD were 90.8 µg/mL (range, 5.7-160 µg/mL) and 45.3 µg/mL (range, 3.2-66.4 µg/mL), respectively, and those data were much higher than the mean of all patients. The mean values of FDP and DD in four patients who died were 244.6 µg/mL (range, 158.4-420 µg/mL) and 102.8 µg/mL (range, 32.8-240 µg/mL), respectively. Both FDP and DD in patients who died were remarkably elevated immediately after trauma. CONCLUSION: FDP and DD levels may be useful parameters for initial evaluation of the severity of trauma, massive blood transfusion, and mortality.

The significance of albumin corrected anion gap in patients with cardiopulmonary arrest.

PURPOSE: The reliable parameter, which can be obtained easily and quickly, is necessary to predict the return of spontaneous circulation (ROSC) of patients with cardiopulmonary arrest (CPA) in the emergency situation. In this study, we evaluated the significance of albumin corrected anion gap (ACAG) for the prediction of ROSC in patients with CPA. PATIENTS AND METHODS: In 166 patients with CPA between January 2009 and December 2010, 132 patients could be analyzed retrospectively. We compared acute physiology and chronic health evaluation (APACHE) II score, sequential organ failure assessment(SOFA) score, anion gap (AG) and ACAG levels between patients with/without ROSC and evaluated the significance of AG and ACAG to predict ROSC in patients with CPA. RESULTS: Both AG and ACAG were significantly lower in patients with ROSC than in patients without ROSC. Both AG and ACAG had the relation with APACHE II and SOFA scores, however, coefficients of correlation with APACHE II and SOFA score were higher in ACAG (r = 0.506) than in AG (r = 0.482). The sensitivity, specificity, positive predictive value, and negative predictive value of ACAG for the prediction of ROSC in patients with CPA were better than those of AG. CONCLUSION: Our study shows that both AG and ACAG have the relation with ROSC and ACAG is better to predict the ROSC following CPR in patients with CPA compared with AG. ACAG can be easily obtained in the emergency situation, and ACAG is a useful parameter to predict ROSC in patients with CPA.

Construction of a high-density reference linkage map of tea (Camellia sinensis).

A few linkage maps of tea have been constructed using pseudo-testcross theory based on dominant marker systems. However, dominant markers are not suitable as landmark markers across a wide range of materials. Therefore, we developed co-dominant SSR markers from genomic DNA and ESTs and constructed a reference map using these co-dominant markers as landmarks. A population of 54 F(1) clones derived from reciprocal crosses between 'Sayamakaori' and 'Kana-Ck17' was used for the linkage analysis. Maps of both parents were constructed from the F(1) population that was taken for BC(1) population. The order of most of the dominant markers in the parental maps was consistent. We constructed a core map by merging the linkage data for markers that detected polymorphisms in both parents. The core map contains 15 linkage groups, which corresponds to the basic chromosome number of tea. The total length of the core map is 1218 cM. Here, we present the reference map as a central core map sandwiched between the parental maps for each linkage group; the combined maps contain 441 SSRs, 7 CAPS, 2 STS and 674 RAPDs. This newly constructed linkage map can be used as a basic reference linkage map of tea.

Constructing highly conducting metal-metal bonded solids by electrocrystallization of [Pt(II)2(RCS2)4] (RCS2(-) = dithiocarboxylato, R = methyl or ethyl).

Partially oxidized one-dimensional (1D) Pt-Pt chain compounds [Pt2(MeCS2)4]4ClO4.5PhCN (1) and [Pt2(EtCS2)4]5(ClO4)2 (2) were synthesized by electrocrystallization of diplatinum(II,II) complexes from different solvents. 1 and 2 consist of 1D Pt-Pt chains of stacked Pt-Pt dimers with short interdimer S...S contacts. Depending on the number of ClO4- per dimer and their positions, 1 forms a regular stack of Pt-Pt dimers, whereas 2 forms pentamer of dimers in the 1D chain. 1 exhibits high electrical conductivity (4.2-8.0 S cm-1) at 300 K and metallic behavior above 125 K. 2 is a semiconductor. 1 exhibits almost temperature independent magnetic susceptibility (ca. 1.1 x 10-4 emu mol-1) which is attributed to Pauli paramagnetism, whereas the spin degree of freedom in 2 has been lost. Although the basic structures are closely related, they exhibited different solid-state properties that depend on the valence state of the platinum atoms and the periodicity within the 1D chain.

Platelets are novel regulators of neovascularization and luteinization during human corpus luteum formation.

The human corpus luteum is a unique endocrine organ that is periodically constructed from the ovulated follicle. During human corpus luteum formation, which is well known as a pathophysiological model for tissue remodeling, the precise mechanisms by which centripetal vascular development is regulated remain unknown. Recently platelets were reported to contain chemoattractive substances with the potential to induce endothelial migration. In this study, we examined the involvement of platelets in the early tissue remodeling process of the human corpus luteum. An immunohistochemical study demonstrated that considerable amounts of red blood cells and CD41-positive platelets were localized at extravascular sites among luteinizing granulosa cells after ovulation. Platelet deposition gradually decreased and became limited near the central cavity toward which microvessels were extending. Platelets were hardly observed in the midluteal phase when the vascular network had already been established. These platelets expressed CD62P/P-selectin and were colocalized with extracellular matrix, suggesting that platelets had been activated by the extracellular matrix. Progesterone production by luteinizing granulosa cells that were isolated from patients undergoing in vitro fertilization therapy was significantly promoted by direct contact with platelets during 4-d culture. Platelet-derived soluble factors induced spreading in granulosa cell morphology. These factors also increased the migration of human umbilical vein endothelial cells, whereas luteinizing granulosa cells attenuated platelet-induced endothelial cell migration. These findings lead us to propose the novel concept that platelets are regulators of endothelial cell migration and granulosa cell luteinization in the remodeling process of the human corpus luteum.

New regulatory mechanisms for human extravillous trophoblast invasion.

Human extravillous trophoblasts (EVT) invade maternal deciduas and reconstructed maternal spiral arteries during early placentation. However, the precise regulatory mechanisms to induce EVT invasion toward arteries and/or to protect EVT from further invasion have not been well understood. Recently, it was found that EVT that had already ceased their invasion, specifically expressed cluster of differentiation (CD9) and dipeptidyl peptidase IV (DPPIV) on their cell surface. In addition, EVT migrating to maternal spiral arteries expressed CC chemokine receptor type-1 (CCR-1), which is a chemokine receptor for regulated on activation normal T cell expressed and secreted (RANTES) and so on. CD9 is associated with integrin molecules on the cell surface and is considered to modulate integrin function. In contrast, DPPIV is a cell surface peptidase that can metabolize RANTES at extracellular sites before its accessing to the chemokine receptors. In vitro functional assay showed that CD9, DPPIV and RANTES are involved in the regulation for EVT invasion. From these findings, it can be proposed that CD9 and DPPIV, including chemokines, are new regulatory factors for human extravillous trophoblasts. (Reprod Med Biol 2005; 4: 189-195).