Mobile and accurate detection system for infection by the 2009 pandemic influenza A (H1N1) virus with a pocket-warmer reverse-transcriptase loop-mediated isothermal amplification.

The 2009 pandemic H1N1 influenza A virus spread quickly worldwide in 2009. Since most of the fatal cases were reported in developing countries, rapid and accurate diagnosis methods that are usable in poorly equipped laboratories are necessary. In this study, a mobile detection system for the 2009 H1N1 influenza A virus was developed using a reverse-transcriptase loop-mediated isothermal amplification (RT-LAMP) kit with a disposable pocket-warmer as a heating device (designated as pwRT-LAMP). The pwRT-LAMP can detect as few as 100 copies of the virus--which is nearly as sensitive as real-time reverse-transcription polymerase chain reaction (RT-PCR)--and does not cross-react with RNA of seasonal influenza viruses. To evaluate the usefulness of the pwRT-LAMP system, nasal swab samples were collected from 56 patients with flu-like symptoms and were tested. Real-time RT-PCR confirmed that the 2009 H1N1 influenza A virus was present in 27 of the 56 samples. Of these 27 positive samples, QuickVue Influenza A+B immunochromatography detected the virus in only 11 samples (11/27; 40.7%), whereas the pwRT-LAMP system detected the virus in 26 of the 56 samples (26/27 of the positive samples; 96.3%). These findings indicate that the mobile pwRT-LAMP system is an accurate diagnostic system for the 2009 H1N1 influenza A virus, and has great potential utility in diagnosing future influenza pandemics.

Stem cells isolated from human dental follicles have osteogenic potential.

OBJECTIVE: Stem cells isolated from human dental follicles as a potential cell source for bone-tissue engineering were examined for correcting a critical bone defect. STUDY DESIGN: Impacted third molars were collected and single cell-derived cell populations were cultivated in growth medium. Single cell-derived cell lines were examined in terms of cell shape, gene expression patterns, differentiation capacity in vitro, and osteogenic potential in vivo. RESULTS: Three distinct cell populations were identified with different morphologies, patterns of gene expression, and differentiation capacity. All 3 cell populations promoted bone formation when transplanted into surgically created critical-size defects in immunodeficient rat calvaria, compared with control animals without cell transplantation, although one of these populations showed a weak capacity for osteogenetic differentiation in vitro. CONCLUSIONS: Human dental follicle can derive at least 3 unique cell populations in culture, all of which promote bone formation in vivo.

Osteogenic differentiation capacity of porcine dental follicle progenitor cells.

This study examined the effect of extracellular matrix (ECM) on the osteogenic differentiation capacity and osteogenesis of dental follicle cells. Single cell-derived porcine dental follicle cells (DFC-I) obtained at the early stage of crown formation in tooth were subcultured and characterized using periodontal ligament cells (PDLC) and bone marrow-derived mesenchymal stem cells (BMSC) as comparison cell populations. The effect of ECM constituents including collagen type I, fibronectin, laminin, and collagen type IV on the differentiation of DFC-1 into osteogenic-lineage cells was evaluated in vitro. In addition, the DFC-1, PDLC, and BMSC populations were compared for osteogenic capacity in vitro by Alizarin red staining and in vivo by transplantation. DFC-I showed different features from PDLC and BMSC. Different components of ECM had different effects on the differentiation of DFC-1 into osteogenic-lineage cells in vitro. Alkaline phosphatase activity and matrix mineralization as early- and late-stage markers of osteogenesis, respectively, supported the differentiation of DFC-1 into osteogenic-related cells in vitro. All three cell types showed equivalent osteogenic capacity in vivo at 4 weeks postoperatively. There were no statistically significant differences among the cell populations with respect to capacity for bone formation. These results suggest a potential application for dental follicle cells in bone-tissue engineering.

Characteristic change and loss of in vivo osteogenic abilities of human bone marrow stromal cells during passage.

Although human bone marrow stromal cells (BMSCs) have the ability to form bone when transplanted, the responsible factors for in vivo osteogenic abilities are poorly understood. Here we report conditions that are required for human BMSCs to demonstrate their in vivo osteogenic abilities. BMSCs were obtained from healthy donors and their in vivo osteogenic abilities were analyzed. Transplantation analyses revealed that the passage number and length of osteogenic induction significantly affected ectopic bone formation. Although 2-week induction increased the percentage of success in bone formation compared with the 1-week induction, BMSCs completely lost their in vivo osteogenic ability after passage 4 regardless of the length of osteogenic induction. Despite their in vivo osteogenic ability, no significant difference was observed in alkaline phosphatase activity or gene expression of osteogenic markers between BMSCs at passages 1 and 3. Differences were only observed in in vitro mineralizing abilities. Application of basic fibroblast growth factor helped to maintain the BMSCs in vivo osteogenic ability; basic fibroblast growth factor altered cell growth and expression of HLA-DR. The results strongly suggest that there are several required conditions for human BMSCs to demonstrate their bone-forming capabilities, which should be further investigated and considered when designing a protocol for clinical bone tissue engineering.

Feasibility and efficacy of bone tissue engineering using human bone marrow stromal cells cultivated in serum-free conditions.

Current standard techniques for bone tissue engineering utilize ex vivo expanded osteogenic cells. However, ex vivo expansion requires serum, which may hinder clinical applications. Here, we report the feasibility and efficacy of bone tissue engineering with human bone marrow stromal cells (BMSCs) expanded in serum-free conditions. Bone marrow was aspirated from 4 healthy donors and adherent cells were cultured in either serum-free medium (STEMPRO((R)) MSC SFM) or conventional serum-containing medium (alpha-MEM supplemented with 10% serum). Efficacy of expansion was greater in serum-free medium. Phenotypically, serum-free expanded BMSCs were smaller in cell-size and showed expression of CD105(++) and CD146(dim). After osteogenic induction, serum-free expanded BMSCs showed lower alkaline phosphatase activity. However, they showed higher responsiveness to induction. In vivo bone-forming ability was also confirmed. In conclusion, bone tissue engineering with serum-free expanded BMSCs is feasible and as efficient as that obtained with BMSCs expanded in conventional serum-containing medium.

Aldosterone inhibits insulin-induced glucose uptake by degradation of insulin receptor substrate (IRS) 1 and IRS2 via a reactive oxygen species-mediated pathway in 3T3-L1 adipocytes.

Serum aldosterone level is clinically known to correlate with body weight and insulin resistance. Because the underlying molecular mechanism is largely unknown, we examined the effect of aldosterone on insulin-induced metabolic signaling leading to glucose uptake in 3T3-L1 adipocytes. Aldosterone reduced the amounts of insulin receptor substrate (IRS) 1 and IRS2 in a time- and dose-dependent manner. As a result, insulin-induced phosphorylation of Akt-1 and -2, and subsequent uptake of 2-deoxyglucose were decreased. Degradation of IRSs was effectively prevented by a glucocorticoid receptor antagonist and antioxidant N-acetylcysteine, but not by a mineralocorticoid receptor antagonist. Because aldosterone induced phosphorylation of IRS1 at Ser(307), responsible kinases were investigated, and we revealed that rapamycin and BMS345541, but neither SP600125 nor calphostin C, conferred for degradation of IRSs. Although lactacystin prevented the degradation of IRSs, glucose uptake was not preserved. Importantly, sucrose-gradient-sediment intracellular fraction analysis revealed that lactacystin did not effectively restore the reduction of IRS1 in the low-density microsome fraction, important for the transduction of insulin's metabolic signaling. These results indicate that aldosterone deteriorates metabolic action of insulin by facilitating the degradation of IRS1 and IRS2 via glucocorticoid receptor-mediated production of reactive oxygen species, and activation of IkappaB Kinase beta and target of rapamycin complex 1. Thus, aldosterone appears to be a novel key factor in the development of insulin resistance in visceral obesity.

Selective diameter control of single-walled carbon nanotubes in the gas-phase synthesis.

A novel approach for selective diameter control of single-walled carbon nanotubes (SWNTs) is performed in the gas-phase growth using two kinds of carbon sources with different decomposition properties; the one carbon source (1st carbon source) is the organic solvent which is difficult to decompose in the reactor and the another carbon source (2nd carbon source) is facile to decompose. The diameter distributions of SWNTs synthesized with various conditions of the flow rate of the 2nd carbon source were investigated by resonant Raman scattering, optical absorption, and photoluminescence (PL) mapping measurements. It was found that increasing the flow rate of the ethylene tends to decrease the diameter of synthesized SWNTs, probably due to the earlier nucleation of SWNTs induced by the ethylene addition. The controlling the flow rate of the ethylene used as a 2nd carbon source can selectively tune the diameter distribution of SWNTs in our growth system.

IR-extended photoluminescence mapping of single-wall and double-wall carbon nanotubes.

A simple and efficient technique is described for measuring photoluminescence (PL) maps of carbon nanotubes (NTs) in the extended IR range (1-2.3 mum). It consists of preparing an NT/surfactant/gelatin film and measuring PL spectra using a combination of a tunable Ti-sapphire laser excitation and FTIR detection. This procedure has been applied to a wide range of single- and double-wall NTs unveiling chirality and diameter distributions that have so far been very difficult to measure. The problems associated with deducing these distributions are discussed by comparing absorption and PL mapping data for NT samples prepared under different conditions.

Supramolecular catalysts for the gas-phase synthesis of single-walled carbon nanotubes.

Reversed micelles containing metallic ions have been used as precursors of novel catalysts for the gas-phase synthesis of single-walled carbon nanotubes (SWNTs). This technique possesses the following advantages: (i) excellent solubility in organic solvents, which are used as reactants and (ii) facile preparation of multicomponent catalysts enabling systematic screening of catalyst compositions for the synthesis of SWNTs. In this study, we report the results of the screening study on the catalytic behavior of Fe-Mo binary catalysts during the synthesis of SWNTs. The results suggested that the catalytic ability was closely related to the strain of the crystal structure of Fe-Mo catalysts formed in the reaction and/or the phase transition caused by dissolution of the Mo atoms. The addition of lithium to the Fe-Mo binary catalysts has revealed an increase in the yield of SWNTs.

Photoluminescence mapping of "as-grown" single-walled carbon nanotubes: a comparison with micelle-encapsulated nanotube solutions.

Band gap photoluminescence (PL) is observed from "as-grown" single-walled carbon nanotubes (SWNTs) in solid form. The relative PL intensities for six specific semiconducting SWNTs are compared directly to those of the micelle-encapsulated SWNTs' solutions to investigate the influence of the micelle dispersion process on PL measurements. The results indicate that sodium dodecyl sulfate (SDS) and sodium cholate (SC) selectively solubilize smaller-diameter nanotubes, whereas sodium dodecylbenzene sulfonate (SDBS) solution does not exhibit significant diameter selectivity within the diameter range studied here (d(t) = 0.829-0.966 nm).

Selective matching of catalyst element and carbon source in single-walled carbon nanotube synthesis on silicon substrates.

We have studied the compatibility of various catalysts for ethylene and ethanol chemical vapor deposition (CVD) syntheses of single-walled carbon nanotubes (SWNTs) on Si substrates. A strong selectivity between the catalyst elemental species and carbon source was found; SWNT yield for Fe (Co) catalysts was much higher for ethylene (ethanol) CVD than for ethanol (ethylene) CVD. This strong and completely opposite selectivity implies significantly different SWNT growth mechanisms for ethanol and ethylene CVD on Si substrates.

Size control of metal nanoparticle catalysts for the gas-phase synthesis of single-walled carbon nanotubes.

Nanoparticle catalysts are essential and indispensable for all syntheses of single-walled carbon nanotubes (SWCNTs). We have prepared size-controlled Co, Co-Mo, and Fe-Mo nanoparticles by the reversed micelle method as the catalysts for the gas-phase pyrolytic synthesis of SWCNTs. From the investigation of the relation between the sizes of the nanoparticles and the alkyl-chain lengths of the cationic surfactants, dialkyldimethylammonium bromides, it has been found that the alkyl groups of the surfactants could play a role in controlling the sizes of the nanoparticles and that the alkyl chain of the surfactant should be preferably less than 10 carbon atoms at most to prepare smaller-size nanoparticles with a narrow size distribution. The reduction of the particle size increases the number of nanoparticles in the colloidal solution and leads to a higher yield of SWCNTs.

[A case report of the 8 year survivor--unresectable liver metastases from advanced gastric cancer (Stage IV) were completely responsive, after 4 years from a total sequential gastrectomy, combining docetaxel treatment to regress the recurrence].

This case was a 69-year-old male who had advanced gastric cancer with unresectable multiple liver metastases (Stage IV). He received a combination therapy consisting of a continuous venous infusion (cisplatin: CDDP 10 mg/body, 5-FU 500 mg/body, day 1-28). As a result, metastatic tumors in the liver completely disappeared and a total gastrectomy was sequentially performed. Four years after the surgery, neck lymph node (LN) metastases and the right adrenal metastasis appeared, and chemotherapy (TS-1, and sequentially TS-1+CDDP) was performed. But, the chemotherapy to eradicate the metastases was hardly enough to be effective. Next, docetaxel (DOC 60 mg/m2 q3w) was started. After 9 courses, they were effective and marked regressions (70%). A total of 15 courses of docetaxel administration were possible until tumor progression recurred. This regimen was not severe in toxicity for the duration except for grade 3 poor appetite. Docetaxel will be a key drug for the gastric cancer. In case of responding well to the chemotherapy, we can hope for an extended long-term survival with a continuation of this regimen.

[A report of two cases--two patients of the extremely advanced hepatocellular carcinoma have responded completely for a long time after a combination therapy consisting of arterial chemotherapy and injection of interferon-alpha].

The prognosis of advanced hepatocellular carcinoma (HCC) is extremely poor. Patient 1 was a 43-year-old male with major portal tumor thrombi. He received combination therapy consisting of continuous arterial infusion (MTX 30 mg/m2, day 1, CDDP/5-FU 6 mg/m2: 250 mg/m2, day 1-14) and subcutaneous injection of IFN-alpha (500 x 10(4) U, 3 times a week, 4 weeks). Patient 2 was a 66-year-old male with major hepatic venous tumor thrombi. He received combination therapy consisting of continuous arterial infusion (5-FU 6 mg/m2: 250 mg/m2, day 1-14) and subcutaneous injection of IFN-alpha (500 x 10(4) U, 3 times a week, 4 weeks). Decrease in tumor was observed in both patients markers and marked regression of tumor was observed in both patients. They are still in complete response. This combination therapy is an effective strategy for advanced HCC.

[Long-term survival after neck lymph node metastases from sigmoid colon cancer--a case report].

We report a 73-year-old female who had been operated on for sigmoid colon cancer and neck lymph node metastases. Tumor size was controlled by systemic chemotherapy for about 3 years. During the chemotherapy, toxicity over grade 2 was not observed and good QOL was maintained on an outpatient basis. It is suggested that low-dose systemic chemotherapy may be useful for patients with poor general status.

Synthesis of crystalline boron nanowires by laser ablation.

Crystalline boron nanowires with tetragonal structure have been synthesized based on laser ablation of a B/NiCo target; the nanowires are sometimes single crystals and have a droplet at one end of the nanowire; the droplet contains B, Ni and Co elements, which indicates that the vapor-liquid-solid (VLS) mechanism may play a key role in the growth of the boron nanowires.