Variable Virulence and Efficacy of BCG Vaccine Strains in Mice and Correlation With Genome Polymorphisms.

Bacille Calmette-Guérin (BCG), an attenuated strain of Mycobacterium bovis, is the only vaccine available for tuberculosis (TB) control. However, BCG is not an ideal vaccine and has two major limitations: BCG exhibits highly variable effectiveness against the development of TB both in pediatric and adult populations and can cause disseminated BCG disease in immunocompromised individuals. BCG comprises a number of substrains that are genetically distinct. Whether and how these genetic differences affect BCG efficacy remains largely unknown. In this study, we performed comparative analyses of the virulence and efficacy of 13 BCG strains, representing different genetic lineages, in SCID and BALB/c mice. Our results show that BCG strains of the DU2 group IV (BCG-Phipps, BCG-Frappier, BCG-Pasteur, and BCG-Tice) exhibit the highest levels of virulence, and BCG strains of the DU2 group II (BCG-Sweden, BCG-Birkhaug) are among the least virulent group. These distinct levels of virulence may be explained by strain-specific duplications and deletions of genomic DNA. There appears to be a general trend that more virulent BCG strains are also more effective in protection against Mycobacterium tuberculosis challenge. Our findings have important implications for current BCG vaccine programs and for future TB vaccine development.

Highly integrated synthesis of heterogeneous nanostructures on nanowire heater array.

We have proposed a new method for the multiplexed synthesis of heterogeneous nanostructures using a top-down fabricated nanowire heater array. Hydrothermally synthesized nanostructures can be grown only on the heated nanowire through nanoscale temperature control using a Joule heated nanowire. We have demonstrated the selective synthesis of zinc oxide (ZnO) nanowires and copper oxide (CuO) nanostructures, as well as their surface modification with noble metal nanoparticles, using a nanowire heater array. Furthermore, we could fabricate an array of heterogeneous nanostructures via Joule heating of individual nanowire heaters and changing of the precursor solutions in a sequential manner. We have formed a parallel array of palladium (Pd) coated ZnO nanowires and gold (Au) coated ZnO nanowires, as well as a parallel array of ZnO nanowires and CuO nanospikes, in the microscale region by using the developed method.

A self-heated silicon nanowire array: selective surface modification with catalytic nanoparticles by nanoscale Joule heating and its gas sensing applications.

We demonstrated novel methods for selective surface modification of silicon nanowire (SiNW) devices with catalytic metal nanoparticles by nanoscale Joule heating and local chemical reaction. The Joule heating of a SiNW generated a localized heat along the SiNW and produced endothermic reactions such as hydrothermal synthesis of nanoparticles or thermal decomposition of polymer thin films. In the first method, palladium (Pd) nanoparticles could be selectively synthesized and directly coated on a SiNW by the reduction of the Pd precursor via Joule heating of the SiNW. In the second method, a sequential process composed of thermal decomposition of a polymer, evaporation of a Pd thin film, and a lift-off process was utilized. The selective decoration of Pd nanoparticles on SiNW was successfully accomplished by using both methods. Finally, we demonstrated the applications of SiNWs decorated with Pd nanoparticles as hydrogen detectors. We also investigated the effect of self-heating of the SiNW sensor on its sensing performance.

Stem cell dynamics in an experimental model of stroke.

We investigated the migration of endogenous neural stem cells (NSCs) toward an infarct lesion in a photo-thrombotic stroke model. The lesions produced by using rose bengal dye (20 mg/kg) with cold light in the motor cortex of Sprague-Dawley rats were also evaluated with sequential magnetic resonance imaging (MRI) from 30 minutes through 8 weeks. Migration of NSCs was identified by immunohistochemistry for nestin monoclonal antibody in the lesion cortex, subventricular zone (SVZ), and corpus callosum (CC). The contrast to noncontrast ratio (CNR) on MRI was greatest at 12 hours in DWI and decreased over time. By contrast, T1-weighted and T2-weighted images showed a constant CNR from the beginning through 8 weeks. MRI of the lesional cortex correlated with histopathologic findings, which could be divided into three stages: acute (edema and necrosis) within 24 hours, subacute (acute and chronic inflammatory cell infiltration) at 2 to 7 days, and chronic (gliofibrosis) at 2 to 4 weeks. The volume of the infarct was significantly reduced by reparative gliofibrosis. The number of nestin(+) NSCs in the contralateral SVZ was similar to that of the ipsilateral SVZ in each group. However, the number of nestin(+) NSCs in the ipsilateral cortex and CC increased at 12 hours to 3 days compared with the contralateral side (p<0.01) and was reduced significantly by 7 days (p<0.01). Active emigration of internal NSCs from the SVZ toward the infarct lesion may also contribute to decreased volume of the infarct lesion, but the self-repair mechanism by endogenous NSCs is insufficient to treat stroke causing extensive neuronal death. Further studies should be focused on amplification technologies of NSCs to enhance the collection of endogenous or transplanted NSCs for the treatment of stroke.

Neuroprotective effects of (-)-epigallocatechin-3-gallate against quinolinic acid-induced excitotoxicity via PI3K pathway and NO inhibition.

Excessive stimulation of the NMDA receptor induces neuronal cell death and is implicated in the development of several neurodegenerative diseases. While EGCG suppresses apoptosis induced by NMDA receptor-mediated excitotoxicity, the mechanisms underlying this process have yet to be completely determined. This study was designed to investigate whether (-)-epigallocatechin-3-gallate (EGCG) plays a neuroprotective role by inhibiting nitric oxide (NO) production and activating cellular signaling mechanisms including MAP kinase, PI3K, and GSK-3beta and acting on the antiapoptotic and the proapoptotic genes in N18D3 neural cells. The cells were pretreated with EGCG for 2 h and then exposed to quinolinic acid (QUIN), a NMDA receptor agonist, 30 mM for 24 h. MTT assay and DAPI staining were used to identify cell viability and apoptosis, respectively, and demonstrated that EGCG significantly increased cell viability and protected the cells from apoptotic death. In addition, EGCG had a capacity to reduce QUIN-induced excitotoxic cell death not only by blocking increase of intracellular calcium levels but also by inhibiting NO production. Gene expression analysis revealed that EGCG prevented the QUIN-induced expression of the proapoptotic gene, caspase-9, and increased that of the antiapoptotic genes, Bcl-XL, Bcl-2, and Bcl-w. Further examination about potential cell signaling candidate involved in this neuroprotective effect showed that immunoreacitivity of PI3K was significantly increased in the cells treated with EGCG. These results suggest that the neuroprotective mechanism of EGCG against QUIN-induced excitotoxic cell death includes regulation of PI3K and modulation of cell survival and death genes through decreasing of intracellular calcium levels and controlling of NO production.

Mass producible and biocompatible microneedle patch and functional verification of its usefulness for transdermal drug delivery.

The key issues in the development of a microneedle patch as a tool for transdermal drug delivery are safety and delivery performance in addition to economical production. In this paper, novel fabrication methods for an inexpensive microneedle patch made of biocompatible polymer are reported, along with functional verifications for the fabricated microneedle patch through animal models. We combined the merits of in-line microneedles, i.e., easy and economical production, with the superior performance of two-dimensionally arrayed microneedles. One-dimensionally fabricated microneedles were assembled to make two-dimensionally arrayed patches to attain our goal. First, we fabricated strips with one-dimensionally arrayed microneedles through deep X-ray lithography on polymethylmethacrylate or another negative photoresist, SU-8, with sharply reduced exposure time. Second, we assembled microneedle strips to make two-dimensionally arrayed microneedles, which we utilized further for fabrication of molding masters. Finally, we prepared microneedle patches made of polycarbonate by hot embossing with these masters. We then demonstrated the actual delivery of exogenous materials through application on skin via animal experiments, and we found no detectable side effects such as inflammation or allergic reactions at the site of application.

[C1772T polymorphism of hypoxia-inducible factor-1 alpha gene in Han ethnic population from Northern China].

OBJECTIVE: To analyze the distribution of single nucleotide polymorphism C1772T (SNP/C1772T) in hypoxia-inducible factor -1 alpha (HIF-1 alpha) gene in Han ethnic population from Northern China. METHODS: Allele frequencies in a sample of 206 healthy Chinese Hans were determined by polymerase chain reaction followed by restriction fragment length polymorphism. RESULT: The genotype frequencies of C1772T observed in Chinese Hans were 93 % and 7% for CC and CT respectively; and allele frequencies were 96% and 4 %for C and T respectively. There was significant difference in distribution of allele frequencies between Chinese Hans and Caucasian. CONCLUSION: The single nucleotide polymorphism of C1772T in HIF-1 alpha gene has difference in different ethnic population.

[The inhibitory effect of Hydrilla verticillata culture water on Microcystic aeruginosa and its mechanism].

This paper studied on the effect and mechanism that the growth of M. aerugonsa was markedly inhibited by the H. verticillata culture water. During treatment, the photosynthetic rate of M. aerugonsa declined, while its respiratory rate and SOD activity increased firstly, then decreased as the treatment went on. Its membrane permeability also increased significantly. TEM photographs showed that the ultrastructure of cell membrane, thylakoid lamella and pith nucleoid of M. aerugonsa were destroyed severely. Inhibitory effects could be observed only when the extracts were extracted by ether. The more extracts from ether, the better inhibitory effect observed. It suggested that the inhibitory effects of H. verticillata on M. aeruginosa were through excreting substances into water. GC/MS analytic result showed that the ether extract mainly consisted of 1,2-benzenedicarboxylic acid diisooctyl ester, dibutyl phthalate, and 1,2-benzenedicarboxylic acid butyl 2-methylpropyl ester.