The purpose of the study is to use O. intermedium DN2 to degrade nicotine in tobacco extracts for making reconstituted tobacco. Firstly, we studied the effects of various factors on degradation of nicotine in the extracts by strain DN2. When we added 0.1% yeast extract into the extracts, adjusted its pH value to 7.0 by ammonia solution, inoculated 15% cultures and maintained fermentation temperature of 30 degrees C, the degradation rate of nicotine by strain DN2 was the fastest. Furthmore, under these conditions, we studied the degradation rates of nicotine in three fed batches culture which carried out in a 30-L reactor, the result showed that the average degradation rate of nicotine by strain DN2 was 140.55 mg/L/h, which was much higher than that reported in other studies. These results indicated that strain DN2 may be useful for reducing nicotine content of reconstituted tobacco.
Nicotine ; metabolism ; Ochrobactrum ; classification ; metabolism ; Plant Extracts ; metabolism ; Tobacco ; chemistry

To improve the extraction of solanesol from tobacco waste, we developed an enzymatic cell wall-breaking process with combined cellulase and ligninase. The effects of reaction time, temperature, pH and enzyme/substrate ratio were determined. The results show that the catalytic effect was better than either single enzyme when the ratio of cellulase to ligninase was 15:1 (U/U). Under the optimized conditions of 175 U/g (enzymes/substrate), tobacco to water 1:5 (W/W), temperature 40 degrees C and pH 6.0, the concentration of solanesol in the solution could reach 0.33 g/L after 8 h. And the average leaching rate reached 96.53% which was 1.68 times of the extraction methods of chemical reflux. It provides new way for the extraction of solanesol from tobacco waste, and worthwhile to be further explored.
Cell Wall ; metabolism ; Cellulase ; metabolism ; Oxygenases ; metabolism ; Plant Leaves ; chemistry ; Terpenes ; isolation & purification ; Tobacco ; chemistry

Nicotine is the main component for smoking addiction. It is widely believed that nicotine dependence is heritable. Many studies are committed to study the effects of specific gene polymorphisms connect with nicotine dependence. Release of dopamine has been considered the most important channel for nicotine dependence. This paper provides a review for recent advance in studies on dopamine system related genetic polymorphisms associated with nicotine dependence.
Animals ; Dopamine ; metabolism ; Humans ; Nicotine ; metabolism ; Polymorphism, Genetic ; Tobacco Use Disorder ; genetics ; metabolism

In order to reveal the induced resistance mechanism of tobacco treated with copper solution to potato virus Y-vein necrosis strain (PVY(N)), disease indexes, contents of virus and some physiological and biochemical indexes in tobacco were studied. The results showed that when treated at the copper concentration of 0.8 mg x L(-1), the symptom displayed and vein necrosis on tobacco were postponed, the disease index and content of virus sharply decreased , and the content of chlorophyll a, chlorophyll b and phenylalanine ammonia lyase (PAL) activity remarkably increased. Furthermore, vein necrosis closely linked to contents of total phenol and flavonoid. In this study, the contents of total phenol and flavonoid were promoted when treated with a solution at the copper concentration of 0.8 mg x L(-1). But the contents of total phenol and flavonoid reached to the first peak at the 3rd day after inoculation, and then decreased to the lowest levels which even were lower than those of the control after inoculating PVY(N). Then the contents of total phenol and flavonoid increased slowly from the 6td but still lower than those of the control. The result implied that spraying copper solution might play an important role in induced resistance of tobacco to vein necrosis disease and strengthen the antiviral capability to PVY(N).
Chlorophyll ; metabolism ; Copper ; pharmacology ; Immunity, Innate ; drug effects ; Phenylalanine Ammonia-Lyase ; metabolism ; Potyvirus ; growth & development ; Tobacco ; drug effects ; metabolism ; virology

OBJECTIVETo determine the changes in the airway inflammation-related cytokine/chemokine profiles after exposure to cigarette smoke (CS) and smoking cessation (SC).

METHODSA total of 18 male C57BL/6 mice were equally divided into three groups: CS group, SC group, and normal control group. The airway resistance, lung morphology, and collagen deposition around airways were determined. HE staining and Masson trichrome staining were used for histopathological analysis. The inflammatory cells in bronchoalveolar lavage fluid (BALF) were assessed. The inflammation-associated cytokines were determined using real-time PCR and immunohistochemistry. Expressions of CXCR3 ligands including the CXCL9, CXCL10, CXCL11 and other cytokines in lung tissue and BALF were also analyzed.

RESULTSThe airway resistance significantly increased in both CS group and SC group when compared with the normal control group. Lung pathological scores in both CS group and SC group were also higher than that in the normal control group, while there was no significant difference between the CS group and SC group. Inflammatory cells including the neutrophils, macrophages, and lymphocytes also increased in both the CS group and SC group at both mRNA and protein levels. The mRNA levels of CXCL9, CXCL10, MMP9, and MMP12 were significantly higher in CS group and SC group than those in the normal control group (all P<0.05). The protein expression levels of CXCL9, CXCL10, CXCL11, MMP2, MMP9, MMP12, and TGF-Β1 were significantly higher in CS group and SC group than those in the normal control group (all P<0.05). Compared with the normal control group,the concentrations of CXCL9, CXCL10, CXCL11, IL-8, and TGF-Β1 in the BALF supernatants of the CS group and SC group significantly increased (P<0.05); in addition, the IL-6 and TNF-Α concentrations also increased in the CS group (both P<0.05).

CONCLUSIONSCS exposure triggers inflammatory cell flux and accumulation in the lung parenchyma and BALF. As a consequence, the inflammatory cytokines increase dramatically. After CS, the cytokines/chemokines can decrease, but is still higher than in non-smokers.

Animals ; Chemokines ; metabolism ; Cytokines ; metabolism ; Lung ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Smoking Cessation ; Tobacco Smoke Pollution

Hd-Zip, a unique transcription factor in plant kingdom, influences the growth, development, and secondary metabolism of plants. Hd-zip Ⅳ is thought to play an important role in trichome development of Schizonepeta tenuifolia. This study aims to explore the functions of StHD1 and StHD8 in Hd-zip Ⅳ subfamily in peltate glandular trichome development. To be specific, the expression patterns of the two genes and interaction between the proteins encoded by them were analyzed based on transcriptome sequencing and two-hybrid screening. The subcellular localization was performed and functions of the genes were verified in tobacco and S. tenuifolia. The results showed that StHD1 and StHD8 had high similarity to HD-Zip Ⅳ proteins of other plants and they all had the characteristic conserved domains of HD-Zip Ⅳ subfamily. They were located in the nucleus. The two genes mainly expressed in young tissues and spikes, and StHD1 and StHD8 proteins interacted with each other. The density and length of glandular trichomes increased significantly in tobacco plants with the overexpression of StHD1 and StHD8. Inhibiting the expression of StHD1 and StHD8 by VIGS(virus-induced gene silencing) in S. tenuifolia resulted in the reduction in the density of peltate glandular trichomes, the expression of key genes related to mono-terpene synthesis, and the relative content of limonene and pulegone, the main components of monoterpene. These results suggested that StHD1 and StHD8 of S. tenuifolia formed a complex to regulate glandular trichomes and affect the biosynthesis of monoterpenes.
Trichomes/metabolism* ; Lamiaceae/genetics* ; Tobacco/genetics* ; Monoterpenes/metabolism* ; Cloning, Molecular ; Plant Proteins/metabolism* ; Gene Expression Regulation, Plant

OBJECTIVES: The prevalence of smoking in schizophrenic patients (74-92%) is higher than that of all psychiatric patients (34-54%) or general population (30-35%). This higher smoking Prevalence is demonstrated even after controlling for known confounders, such as marital status, alcohol use, and socioeconomic status. This study was conducted to determine whether there would be any difference in nicotine intake and metabolism between schizophrenics and normal controls. METHODS: Sixteen schizophrenic patients and sixteen normal controls were collected. All subjects were supplied with a pack of cigarette a day. Urinary cotinine excretion was measured by using gas chromatographic mass spectrometric method. RESULTS: Cotinine excretion was significantly increased in schizophrenic patients compared to normal controls (p<0.05). None of variables such as age at initial smoking, the average number of cigarettes at initial smoking, pack year (packs daily smoked x smoking year), abstinence history were found to influence cotinine levels when examined via the ANOVA, even when the interaction with diagnosis was considered. CONCLUSION: This result suggests that nicotine intake and consumption are increased in schizophrenic patients compared to normal controls, which can be an attempt to improve sensory inhibition and counteract neuronal effect of antipsychotic medications.
Cotinine* ; Diagnosis ; Humans ; Marital Status ; Metabolism ; Neurons ; Nicotine ; Prevalence ; Schizophrenia ; Smoke ; Smoking ; Social Class ; Tobacco Products

OBJECTIVE: To assess the potential of transient expression of recombinant human plasminogen activator (rhPA) in plants as a cost-effective approach for recombinant rhPA production. METHODS: Tobacco mosaic virus-based expression vector pTMV rhPA-NSK and plant binary expression vector pJ Zera-rhPA were constructed by sequence synthesis and subcloning. The two vectors were inoculated on either or leaves agroinfiltration. The expression of recombinant rhPA in leaves was examined using Western blotting and ELISA, and the fibrinolysis activity of plant-produced rhPA was assessed by fibrin agarose plate assay (FAPA). RESULTS: Five to nine days after infiltration with an inoculum containing pTMV rhPA-NSK, necrosis appeared in the infiltrated area on the leaves of both plants, but intact recombinant rhPA was still present in the necrotic leaf tissues. The accumulation level of recombinant rhPA in infiltrated leaves was significantly higher than that in leaves ( < 0.05). The yield of recombinant rhPA was up to 0.6% of the total soluble protein (or about 60.0 μg per gram) in the fresh leaf biomass at 7 days post-inoculation. The plant-derived rhPA was bioactive to convert inactive plasminogen to active plasmin. No necrosis occurred in pJ Zera-rhPA-infiltrated leaves. The Zera-rhPA protein was partially cleaved between the site of Zera tag and rhPA sequence in both leaves. We speculated that the formation of Zera tags-induced particles in the plant cells was a dynamic process of progressive aggregation in which some of the soluble polypeptides were encapsulated in these particles. CONCLUSIONS Enzymatically active recombinant rhPA can be rapidly expressed in tobacco plants using the plant viral ampliconbased system, which offers a promising alternative for cost-effective production of recombinant rhPA.
Humans ; Plant Leaves ; Plants, Genetically Modified ; Plasminogen ; Plasminogen Activators ; metabolism ; Recombinant Proteins ; Tobacco

Recombinant proteins provide new means for disease treatment, while creating considerable economic benefits. Using commercial crops (mainly tobacco), cereal crops, legumes, and vegetable crops to produce recombinant proteins with medicinal value is a hot-spot for research in "molecular farming". Although many recombinant proteins have been expressed in plants, only a small number have been successfully put into use. To overcome the problems that greatly hamper the development of recombinant protein production in plants, researchers have improved expression systems to increase the yield of recombinant proteins. Starting from analyzing the problems of low yield and/or low biological activity of recombinant proteins produced by plants, the optimization strategies to solve these problems were reviewed, and future research directions for improving the yield of recombinant proteins produced by plants were proposed.
Crops, Agricultural/genetics* ; Plant Proteins/metabolism* ; Plants, Genetically Modified/genetics* ; Recombinant Proteins ; Tobacco/genetics*

Studies have shown that transgenic plants expressing antiapoptotic genes from baculovirus and animals increase resistance to biotic and abiotic stress. However, the mechanism under these resistances is conjectural, or in some cases even controversy. In the present study, the p35 gene from baculovirus Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) was expressed in tobacco, and for the first time P35 protein was detected in transgenic plants by Western blotting. Inoculation of T1 transgenic tobacco leaves with tobacco mosaic virus (TMV) showed enhanced resistance, and DNA laddering was observed after TMV infection in control but not in transgenic plants. DAB staining showed that TMV infection did not affect peroxide induction of transgenic plants, Western blotting analysis of PR1 protein also showed no difference of control and transgenic plants. Inoculation of fungus (Sclerotinia sclerotiorum) using a detached leaf assay showed enhanced resistance of transgenic leave tissue. RT-PCR analysis demonstrated that p35 gene expression induced earlier expression of PR1 gene after S. sclerotiorum infection. Taken together, our results suggest that the mechanism under enhanced disease resistance by P35 protein is possibly related to the activation of PR-related proteins in addition to the inhibition of programmed cell death, depending on the pathogens challenged.
Gene Expression Regulation, Plant ; Immunity, Innate ; Plant Diseases ; genetics ; Plants, Genetically Modified ; genetics ; immunology ; virology ; Tobacco ; genetics ; immunology ; virology ; Tobacco Mosaic Virus ; Transformation, Genetic ; Viral Proteins ; genetics ; metabolism