Construction and Evaluation of Chloroplast Expression Vectors in Higher Plants

Plastid transformation has a number of advantages in comparison with nuclear transformation. Currently, only tobacco (Nicotiana tabacum) is routinely used in plastid transformation. Here we constructed a series of chloroplast expression vectors specific for spinach (pCEV1), tomato (pCEV2 and pCEV3), and N. benthamiana (pCEV4). Selection marker aminoglycoside 3′-adenyltransferase (aadA) conferring spectinomycin resistance was used in pCEV1, pCEV2, and pCEV4, while selection marker neomycin phosphotransferase II (nptII) was used in pCEV3. The expression cassette in these vectors was integrated in the intergenic spacer between trnI and trnA of plastid genome via homologous recombination. Several transgenes, including a reporter gene encoding GFP:GUS fusion protein and genes from tomato (lycopene b-cyclase, z-carotene desaturase) and bamboo mosaic virus satellite RNA (encoding coat protein CP20), were independently cloned into some of these vectors. Transient GUS expression was detected in spinach leaves bombarded by pCEV1/GFP-GUS. Functional expression of selection markers aadA and nptII was demonstrated for spinach, tomato, and N. benthamiana. Seedling assay from T0 self-pollinated plant of transplastomic N. benthamiana confirmed maternal inheritance of transgenes, and genomic PCR analysis confirmed integration of transgenic expression cassette into the plastid genome of N. benthamiana. Moreover, auxiliary vectors pECaad and pECnpt are also reported

The Effect of Financial Resources on Fertility: Evidence from Administrative Data on Lottery Winners

This paper utilizes wealth shocks from winning lottery prizes to examine the effect of financial resources on fertility. Using administrative data on Taiwanese lottery winners and a difference-in-differences design, we compare the trend in fertility between households receiving lottery prizes of more than 1 million NT$(33,000 US$) with those winning less than 10,000 NT$(330 US$). The results show that the receipt of a big lottery prize significantly increases fertility, and effects are driven by households with less financial resources. Moreover, big lottery wins mainly trigger childless households to have children and induce people to get married earlier

Knockdown of PsbO leads to induction of HydA and production of photobiological H2 in the green alga Chlorella sp. DT

Green algae are able to convert solar energy to H2 via the photosynthetic electron transport pathway under certain conditions. Algal hydrogenase (HydA, encoded by HYDA) is in charge of catalyzing the reaction: 2H+ + 2e− ↔ H2 but usually inhibited by O2, a byproduct of photosynthesis. The aim of this study was to knockdown PsbO (encoded by psbO), a subunit concerned with O2 evolution, so that it would lead to HydA induction. The alga, Chlorella sp. DT, was then transformed with short interference RNA antisense-psbO (siRNA-psbO) fragments. The algal mutants were selected by checking for the existence of siRNA-psbO fragments in their genomes and the low amount of PsbO proteins. The HYDA transcription and the HydA expression were observed in the PsbO-knockdown mutants. Under semi-aerobic condition, PsbO-knockdown mutants could photobiologically produce H2 which increased by as much as 10-fold in comparison to the wild type

Nocturnal CPAP improves walking capacity in COPD patients with obstructive sleep apnoea

BACKGROUND: Exercise limitation is an important issue in patients with chronic obstructive pulmonary disease (COPD), and it often co-exists with obstructive sleep apnoea (overlap syndrome). This study examined the effects of nocturnal continuous positive airway pressure (CPAP) treatment on walking capacity in COPD patients with or without obstructive sleep apnoea. METHODS: Forty-four stable moderate-to-severe COPD patients were recruited and completed this study. They all underwent polysomnography, CPAP titration, accommodation, and treatment with adequate pressure. The incremental shuttle walking test was used to measure walking capacity at baseline and after two nights of CPAP treatment. Urinary catecholamine and heart rate variability were measured before and after CPAP treatment. RESULTS: After two nights of CPAP treatment, the apnoea-hypopnoea index and oxygen desaturation index significantly improved in both overlap syndrome and COPD patients, however these changes were significantly greater in the overlap syndrome than in the COPD group. Sleep architecture and autonomic dysfunction significantly improved in the overlap syndrome group but not in the COPD group. CPAP treatment was associated with an increased walking capacity from baseline from 226.4 ± 95.3 m to 288.6 ± 94.6 m (P < 0.05), and decreased urinary catecholamine levels, pre-exercise heart rate, oxygenation, and Borg scale in the overlap syndrome group. An improvement in the apnoea-hypopnoea index was an independent factor associated with the increase in walking distance (r = 0.564). CONCLUSION: Nocturnal CPAP may improve walking capacity in COPD patients with overlap syndrome. TRIAL REGISTRATION: NCT0091426

Schwann Cell Migration Induced by Earthworm Extract via Activation of PAs and MMP2/9 Mediated through ERK1/2 and p38

The earthworm, which has stasis removal and wound-healing functions, is a widely used Chinese herbal medicine in China. Schwann cell migration is critical for the regeneration of injured nerves. Schwann cells provide an essentially supportive activity for neuron regeneration. However, the molecular migration mechanisms induced by earthworms in Schwann cells remain unclear. Here, we investigate the roles of MAPK (ERK1/2, JNK and p38) pathways for earthworm-induced matrix-degrading proteolytic enzyme (PAs and MMP2/9) production in Schwann cells. Moreover, earthworm induced phosphorylation of ERK1/2 and p38, but not JNK, activate the downstream signaling expression of PAs and MMPs in a time-dependent manner. Earthworm-stimulated ERK1/2 and p38 phosphorylation was attenuated by pretreatment with U0126 and SB203580, resulting in migration and uPA-related signal pathway inhibition. The results were confirmed using small interfering ERK1/2 and p38 RNA. These results demonstrated that earthworms can stimulate Schwann cell migration and up-regulate PAs and MMP2/9 expression mediated through the MAPK pathways, ERK1/2 and p38. Taken together, our data suggests the MAPKs (ERK1/2, p38)-, PAs (uPA, tPA)-, MMP (MMP2, MMP9) signaling pathway of Schwann cells regulated by earthworms might play a major role in Schwann cell migration and nerve regeneration

Levels and values of circulating endothelial progenitor cells, soluble angiogenic factors, and mononuclear cell apoptosis in liver cirrhosis patients

BACKGROUND: The roles of circulating endothelial progenitor cell (EPC) and mononuclear cell apoptosis (MCA) in liver cirrhosis (LC) patients are unknown. Moreover, vascular endothelial growth factor (VEGF) and stromal cell-derived factor (SDF)-1α are powerful endogenous substances enhancing EPC migration into circulation. We assessed the level and function of EPCs [CD31/CD34 (E(1)), KDR/CD34 (E(2)), CXCR4/CD34 (E(3))], levels of MCA, VEGF and SDF-1α in circulation of LC patients. METHODS: Blood sample was prospectively collected once for assessing EPC level and function, MCA, and plasma levels of VEGF and SDF-1α using flow cytometry and enzyme-linked immunosorbent assay (ELISA), respectively, in 78 LC patients and 25 age- and gender-matched healthy controls. RESULTS: Number of EPCs (E(1), E(2), E(3)) was lower (all p < 0.0001), whereas SDF-1α level and MCA were higher (p < 0.001) in study patients compared with healthy controls. Number of EPCs (E(2), E(3)) was higher but MCA was lower (all p < 0.05) in Child's class A compared with Child's class B and C patients, although no difference in VEGF and SDF-1α levels were noted among these patients. Chronic hepatitis B and esophageal varices bleeding were independently, whereas chronic hepatitis C, elevated aspartate aminotransferase (AST), and decompensated LC were inversely and independently correlated with circulating EPC level (all p < 0.03). Additionally, angiogenesis and transwell migratory ability of EPCs were reduced in LC patients than in controls (all p < 0.001). CONCLUSION: The results of this study demonstrated that level, angiogenic capacity, and function of circulating EPCs were significantly reduced, whereas plasma levels of SDF-1α and circulating MCA were substantially enhanced in cirrhotic patients

RSC96 Schwann Cell Proliferation and Survival Induced by Dilong through PI3K/Akt Signaling Mediated by IGF-I

Schwann cell proliferation is critical for the regeneration of injured nerves. Dilongs are widely used in Chinese herbal medicine to remove stasis and stimulate wound-healing functions. Exactly how this Chinese herbal medicine promotes tissue survival remains unclear. The aim of the present study was to investigate the molecular mechanisms by which Dilong promote neuron regeneration. Our results show that treatment with extract of Dilong induces the phosphorylation of the insulin-like growth factor-I (IGF-I)-mediated phosphatidylinositol 3-kinase/serine-threonine kinase (PI3K/Akt) pathway, and activates protein expression of cell nuclear antigen (PCNA) in a time-dependent manner. Cell cycle analysis showed that G1 transits into the S phase in 12–16 h, and S transits into the G2 phase 20 h after exposure to earthworm extract. Strong expression of cyclin D1, cyclin E and cyclin A occurs in a time-dependent manner. Small interfering RNA (siRNA)-mediated knockdown of PI3K significantly reduced PI3K protein expression levels, resulting in Bcl2 survival factor reduction and a marked blockage of G1 to S transition in proliferating cells. These results demonstrate that Dilong promotes the proliferation and survival of RSC96 cells via IGF-I signaling. The mechanism is mainly dependent on the PI3K protein

Cathodoluminescence spectra of gallium nitride nanorods

Gallium nitride [GaN] nanorods grown on a Si(111) substrate at 720°C via plasma-assisted molecular beam epitaxy were studied by field-emission electron microscopy and cathodoluminescence [CL]. The surface topography and optical properties of the GaN nanorod cluster and single GaN nanorod were measured and discussed. The defect-related CL spectra of GaN nanorods and their dependence on temperature were investigated. The CL spectra along the length of the individual GaN nanorod were also studied. The results reveal that the 3.2-eV peak comes from the structural defect at the interface between the GaN nanorod and Si substrate. The surface state emission of the single GaN nanorod is stronger as the diameter of the GaN nanorod becomes smaller due to an increased surface-to-volume ratio

Flux tunable graphene-based superconducting quantum circuits coupled to 3D cavity

Correlation between transmon and its composite Josephson junctions (JJ) plays an important role in designing new types of superconducting qubits based on quantum materials. It is desirable to have a type of device that not only allows exploration for use in quantum information processing but also probing intrinsic properties in the composite JJs. Here, we construct a flux-tunable 3D transmon-type superconducting quantum circuit made of graphene as a proof-of-concept prototype device. This 3D transmon-type device not only enables coupling to 3D cavities for microwave probes but also permits DC transport measurements on the same device, providing useful connections between transmon properties and critical currents associated with JJ's properties. We have demonstrated how flux-modulation in cavity frequency and DC critical current can be correlated under the influence of Fraunhofer pattern of JJs in an asymmetric SQUID. The correlation analysis was further extended to link the flux-modulated transmon properties, such as flux-tunability in qubit and cavity frequencies, with SQUID symmetry analysis based on DC measurements. Our study paves the way towards integrating novel materials for exploration of new types of quantum devices for future technology while probing underlying physics in the composite materials