Nonradioactive Northern blotting with biotinylated and digoxigenin-labeled RNA probes

The application of nonradioactive RNA probes for Northern blotting offers the advantage of a rapid turn-around time for results without the loss of sensitivity for target mRNA detection. However, a problem that has impeded the widespread use of nonradioactive RNA probes for use in Northern blotting is the difficulty in stripping these probes from nylon membranes after hybridization. In this report we describe two protocols for stripping digoxigenin (Dig)-labeled RNA probes from nylon membranes. One protocol utilizes a phosphate-buffered formamide stripping solution to remove nonchemically modified (regular) RNA probes while the other method utilizes strippable probes that were produced with a chemically modified nucleotide (CTP) and removed by a specific stripping solution. This latter method was developed by Ambion Inc. and is called Strip-EZ. We also describe a protocol for the detection of two separate rat mRNAs using both biotin and digoxigenin-labeled RNA probes that does not require stripping the membrane after hybridization. Finally, we describe the use of another new labeling technology, called Chem-Link, that quickly and conveniently labels RNA for use in Northern blotting.     

A new class of tobacco chitinases homologous to bacterial exo-chitinases displays antifungal activity

A novel chitinase gene of tobacco was isolated and characterized by DNA sequence analysis of a genomic clone and a cDNA clone. Comparative sequence analysis of both clones showed an identity of 94%. The proteins encoded by these sequences do not correspond to any of the previously characterized plant chitinases of classes I-IV and are designated as class V chitinases. Comparison of the chitinase class V peptide sequence with sequences in the Swiss Protein databank revealed significant sequence similarity with bacterial exo-chitinases from Bacillus circulans, Serratia marcescens and Streptomyces plicatus. It was demonstrated that class V chitinase gene expression is induced after treatment of tobacco with different forms of stress, like TMV-infection, ethylene treatment, wounding or ultraviolet irradiation. Two related chitinase class V proteins of 41 and 43 kDa were purified from Samsun NN tobacco leaves inoculated with tobacco mosaic virus. The proteins were purified by Chelating Superose chromatography and gel filtration. In vitro assays demonstrated that class V chitinases have endo-chitinase activity and exhibit antifungal activity toward Trichoderma viride and Alternaria radicina. In addition, it was shown that class V chitinase acts synergistically with tobacco class I beta-1,3-glucanase against Fusarium solani germlings.     

Ionic transport studies of the glassy silver vanadomolybdate system

Ionic conductivity of the Ag₂O-MoO₃-V₂0₅ system has been studied over a wide range of frequency, temperature and composition. A narrower glass forming region has been found in comparison to the corresponding Ag₂O-MoO₃-P₂O₅ and Ag₂O-B₂O₃-P₂O₅ systems. The highest conductivity at room temperature, _rt, = 3.21 × 10^–6–1 cm^–1 (d.c.) with an activation energy,E _act, of 0.466 eV, was observed for the glass former's ratio of unity. Further, it reached a maximum value of 2.2 × 10^–2¨-1 cm^–1 withE _act = 0.153 eV when the oxide-base glass was dissolved with Agl. D.c. conductivity, hopping rate and relaxation time in the present system have been found to be characterized by the same activation energy.     

Miscibility of poly(methoxymethyl methacrylate) and poly(methylthiomethyl methacrylate) with poly(vinylidene fluoride)

Summary The miscibility behaviour of poly(methoxymethyl methacrylate) (PMOMA) and poly(methylthiomethyl methacrylate) (PMTMA) with poly(vinylidene fluoride) (PVDF) was examined by differential scanning calorimetry. PMOMA/PVDF blend system was judged to be miscible on the bases of the presence of a single, composition-dependent glass transition for the blend and a pronounced melting point depression of the PVDF component. Furthermore, lower critical solution temperature (LCST) behaviour was observed for all PMOMA/PVDF blends. PMTMA/PVDF blends were found to be immiscible. Based on the melting point depression of PVDF in PMOMA/PVDF blends, the interaction parameter B was found to be -14.5 J/cm³.     

机械总院研发立式冷摆动辗压机填补国内空白

机械科学研究总院北京机电研究所承担“2000KN立式冷摆动辗压机开发研究”项目通过验收,研发的2000kN矿式冷摆动辗压机具有自主知识产权,填补了国内空白,达到国际先进水平,人选“2013度中关村科技园区首台（套）重大技术装备试验、示范项目。     

Emergence of photoluminescence on bulk MoS<Subscript>2</Subscript> by laser thinning and gold particle decoration

We demonstrate a facile and effective approach to significantly improve the photoluminescence of bulk MoS₂ via laser thinning followed by gold particle decoration. Upon laser thinning of exfoliated bulk MoS₂, photoluminescence emerges from the laser-thinned region. After further treatment with an AuCl₃ solution, gold particles self-assemble on the laser-thinned region and thick edges, further increasing the fluorescence of bulk MoS₂ 28 times and the Raman response 3 times. Such fluorescence enhancement can be attributed to both surface plasmon resonance and p-type doping induced by gold particles. The combination of laser thinning and AuCl₃ treatment enables the functionalization of bulk MoS₂ for optoelectronic applications. It can also provide a viable strategy for mask-free and area-selective p-type doping on single MoS₂ flakes.

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Prevalence and incidence of hand dermatitis in hairdressing apprentices: results of the POSH study. Prevention of occupational skin disease in hairdressers

OBJECTIVE: Occupational skin changes in hairdressers are very common. Morbidity, however, has not yet been quantified precisely. METHODS: A cohort of 2,352 hairdressing apprentices (of the 2,570 invited to participate, i.e., 91.5% response) was prospectively followed for the duration of their vocational training (3 years) by three examinations. Three waves (years) were recruited in 1992, 1993, and 1994 from 15 vocational training schools in northwestern Germany. RESULTS: The point prevalence of (mostly slight) irritant skin changes of the hands increased from 35.4% in the initial examination to 47.5% in the intermediate examination and to 55.1% in the final examination. Given a more conservative definition of a case of "hand dermatitis," these estimates were 12.9%, 23.5%, and 23.9%, respectively. Altogether, 34.3 and 15.2 cases of "skin changes (any degree)" and "hand dermatitis," respectively, in 100 person-years were observed during the study period. The incidence rate, i.e., the number of newly diseased study participants in relation to the person-time at risk contributed, decreased in the course of the study. The proportion of dropouts until final follow-up was 51.8%. CONCLUSION: The present results appear to lie in a range with those found in other, much smaller cohort studies. However, comparison of the results is hampered either by the lack of a clear definition or by a different definition of "person-time at risk" or "a case of hand dermatitis." As compared with an external control group of office apprentices, the incidence was several times higher in hairdressing apprentices, which points to the high risk for skin damage in this occupation.     

Assessing patients'' priorities and perceptions of the quality of health care: the development of the QUOTE-Rheumatic-Patients instrument

BACKGROUND: Stress can exacerbate a number of psychiatric disorders, many of which are associated with prefrontal cortical (PFC) cognitive deficits. Biochemical studies demonstrate that mild stress preferentially increases dopamine turnover in the PFC. Our study examined the effects of acute, mild stress exposure on higher cognitive function in monkeys and the role of dopaminergic mechanisms in the stress response. METHODS: The effects of loud (105-dB) noise stress were examined on a spatial working memory task (delayed response) dependent on the PFC, and on a reference memory task with similar motor and motivational demands (visual pattern discrimination) dependent on the inferior temporal cortex. The role of dopamine mechanisms was tested by challenging the stress response with agents that decrease dopamine receptor stimulation. RESULTS: Exposure to noise stress significantly impaired delayed-response performance. Stress did not impair performance on "0-second" delay control trials and did not alter visual pattern discrimination performance, which is consistent with impaired PFC cognitive function rather than nonspecific changes in performance. Stress-induced deficits in delayed-response performance were ameliorated by pretreatment with drugs that block dopamine receptors (haloperidol, SCH 23390) or reduce stress-induced PFC dopamine turnover in rodents (clonidine, naloxone hydrochloride). CONCLUSIONS: These results indicate that stress impairs PFC cognitive function through a hyperdopaminergic mechanism. Stress may take the PFC "off-line" to allow more habitual responses mediated by posterior cortical and subcortical structures to regulate behavior. This mechanism may have survival value, but may often be maladaptive in human society, contributing to the vulnerability of the PFC in many neuropsychiatric disorders.     

FRC measurement in intensive care patients. A definition of standards

Determination of Functional Residual Capacity (FRC) can be performed through washout methods, indicator gas dilution or bodyplethysmography. Some of these techniques have been adapted for use in intensive care patients whilst being mechanically ventilated. However, most measurement setups are bulky, cumbersome to use and their running costs are high. Hence FRC measurement has not become a routine method in intensive care although it offers considerable advantages in the management of ventilated patients such as the determination of "best PEEP", the detection of progressive alveolar collapse in the course of acute lung injury and during weaning from mechanical ventilation. Up to now most efforts to improve and simplify FRC measurement were made at the expense of accuracy. An ideal method ought to be accurate, easy to handle and cost-effective. It should supply not only FRC data but also information about intrapulmonary gas distribution and dead space. These demands can be met using modern data acquisition software. The pros and cons of all methods available for FRC measurement are discussed in view of their suitability for intensive care patients. A conventional nitrogen washout using emission spectroscopy for measurement of nitrogen concentration gives satisfying exact values for the determination of the parameters mentioned above. The measurement error can be lowered under 5% by special corrections for flow and nitrogen signal (delay and rise times, changes of gas viscosity). For flow measurement a normal pneumotachograph can be used. Using a laptop computer for data acquisition the bed-side monitor fulfills most of the demands in intensive care. It is then also possible to measure indices of intrapulmonary gas distribution such as Alveolar Mixing Efficiency and Lung Clearance Index.