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Introduction: The COVID-19 pandemic created challenges for forward-deployed military units to Western Africa. Austere military environments afford multiple avenues to transmit COVID-19 amongst service members., Materials and Methods: A COVID-19 outbreak on a military base in Western Africa spanning over 100 days is statistically analyzed using a Pearson's correlation coefficient. Furthermore, a COVID-19 reproductive number (R0) is evaluated to examine the relationship between specific command-directed policies to mitigate COVID-19 transmission., Results: The multidisciplinary partnership of military command, medical, and public health leadership implemented evidence-based and epidemiologically informed COVID-19 preventive base-wide policies, including appropriate isolation/quarantine policies. The R0 for the outbreak was 0.03 and remained <1 for the outbreak duration. This base remained COVID-19 free for multiple weeks after policy implementation., Conclusions: The implementation of practical mitigating base-wide policies through seamless communication between military command/medical/public health leadership resolved the COVID-19 outbreak while maintaining mission readiness. Weekly COVID-19 testing epidemiological data may be utilized by commanders to direct further decision-making on tightening/loosening base-wide policy restrictions for continued mission-essential operations, e.g., security, food service, or airfield operations., (© The Association of Military Surgeons of the United States 2022. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Aims: Investigation of the influence of cultivation time and sea buckthorn press cake (Hippophaë rhamnoides) dosage on mycelium yield of Inonotus obliquus in submerged cultivation and on the yield, monomer composition, and macromolecular properties of the exopolysaccharides (EPS) from culture media and intracellular polysaccharides (IPS) extracted from mycelia., Methods and Results: Supplementation at 5 g l -1 combined with cultivation time of 250 h granted highest yield increase in mycelia (by 122%). The supplementation reduced extraction yield and decreased the molecular weight of the main IPS population. The supplementation increased production and molecular weight of EPS. The relative content of arabinose and rhamnose in EPS positively correlated with dosage of the press cake. The press cake supplementation increased the content of galacturonic acid in IPS, but not in EPS., Conclusion: Sea buckthorn press cake is a food industry fibrous side stream with high oil content. It increases the cultivation yield of Inonotus obliquus mycelium and influences the produced polysaccharides., Significance and Impact of the Study: Mycelium is a resource of bioactive polysaccharides, attracting the interest of nutraceutical companies. Sea buckthorn press cake is a promising supplement for increasing mycelium production. The utilization of this agricultural side stream would therefore favour circular economy., (© 2021 The Authors. Journal of Applied Microbiology published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology.)

This comment piece explores the decision in Whittington Hospital NHS Trust v XX [2020] UKSC 14. It argues that despite notable shifts in public policy in respect of the acceptability of surrogacy as a means of family formation in the past twenty years, the Supreme Court has taken a step too far in deciding that foreign commercial surrogacy is as widely socially accepted. This impacts on the reasonableness of any claim for damages in negligence for the costs of commercial surrogacy. It is posited that the issue of whether damages for foreign commercial surrogacy are reasonable or not will be the key battleground in future negligence cases of this type., (© The Author(s) 2020. Published by Oxford University Press; All rights reserved. For permissions, please email: journals.permissions@oup.com.)

The Asian citrus psyllid, Diaphorina citri Kuwayama, is the most serious pest of citrus because it is a vector for the highly destructive citrus greening disease (huanglongbing, HLB). Currently, insecticide applications are being used widely to control psyllid populations, thereby suppressing the spread of HLB. In the present study, topical application bioassays were performed to detect the joint actions of beta-cyfluthrin and thiamethoxam or tolfenpyrad against D. citri adults in the laboratory. In 2019, a field plot experiment was conducted to evaluate the control efficacies of beta-cyfluthrin+thiamethoxam 22% capsule suspension and beta-cyfluthrin+tolfenpyrad 30% microemulsion against D. citri using foliar sprays. For the former, a 9:13 mass ratio had the highest synergistic effect, with a cotoxicity coefficient of 188.64. For the latter, a 5:25 mass ratio had the highest synergistic effect, with a cotoxicity coefficient of 153.94. A one-time foliar spray of the former at 30-40 mg/kg or of the latter at 40-60 mg/kg effectively controlled D. citri, with control efficacies varying from 80.1 to 99.4% or 80.4 to 100.0%, during the 3-30 d after treatment, respectively. Moreover, field observations indicated that these foliar sprays at the tested rates had no negative effects on citrus trees. Thus, foliar sprays of beta-cyfluthrin+thiamethoxam or beta-cyfluthrin+tolfenpyrad under the given conditions may control D. citri., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Whitefly-transmitted begomoviruses are economically important plant pathogens that cause severe problems in many crop plants, such as tomato, papaya, cotton, and tobacco. Tomato yellow leaf curl virus (TYLCV) is a typical monopartite begomovirus that has been extensively studied, but methods that can efficiently control begomoviruses are still scarce. In this study, we combined artificial microRNA (amiRNA)-mediated silencing technology and clay nanosheet-mediated delivery by spraying and developed a method for efficiently preventing TYLCV infection in tomato plants. We designed three amiRNAs that target different regions of TYLCV to silence virus-produced transcripts. Three plant expression vectors expressing pre-amiRNAs were constructed, and recombinant plasmid DNAs (pDNAs) were loaded onto nontoxic and degradable layered double hydroxide (LDH) clay nanosheets. LDH nanosheets containing multiple pDNAs were sprayed onto plant leaves. We found that the designed amiRNAs were significantly accumulated in leaves 7 days after spraying, while the pDNAs were sustainably detected for 35 days after the spray, suggesting that the LDH nanosheets released pDNAs in a sustained manner, protected pDNAs from degradation and efficiently delivered pDNAs into plant cells. Importantly, when the LDH nanosheets coated with pDNAs were sprayed onto plants infected by TYLCV, both the disease severity and TYLCV viral concentration in sprayed plants were significantly decreased during the 35 days, while the levels of H 2 O 2 were significantly increased in those plants. Taken together, these results indicate that LDH nanosheets loaded with pDNAs expressing amiRNAs can be a sustainable and promising tool for begomovirus control.

Diaphania hyalinata (Linnaeus, 1767) is one of the main pests of the cucurbit crops. Biological control and botanicals are used in integrated pest management (IPM), especially in agro-ecological agricultures where the use of synthetic pesticides is restricted. Toxicological effects of plant essential oils on target and nontarget organisms should be evaluated to determine its use in IPM. The toxicity of ginger, peppermint, oregano, and thyme essential oils to D. hyalinata eggs, larvae, and pupae and their selectivity to the parasitoid Trichospilus pupivorus and the LC50, LC99 and the relative selectivity index (RSI) of these botanicals were determined. The eggs of D. hyalinata were more sensitive to the botanicals applied than its larvae and pupae, with higher toxicity of oregano and thyme essential oils, followed by those of peppermint and ginger. Topical application was the least toxic method to D. hyalinata larvae. Ginger, peppermint, and thyme essential oils were more toxic by ingestion and oregano by contact to D. hyalinata larvae. The essential oil concentrations applied to D. hyalinata pupae prevent the emergence of adults of this insect with the oregano essential oil showing the greatest toxicity. Peppermint, ginger, thyme, and oregano essential oils were selective to T. pupivorus with RSI50 of 5.40, 1.38, 8.15, and 6.98 and RSI99 of 1.54, 2.53, 3.90, and 4.16 respectively. The ginger, peppermint, oregano, and thyme essential oils were toxic to immature D. hyalinata and selective to T. pupivorus females presenting potential as an alternative control in the IPM of this pest in Cucurbitaceae crops., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The function of the transcription factor KNOTTED ARABIDOPSIS THALIANA7 (KNAT7) is still unclear since it appears to be either a negative or a positive regulator for secondary cell wall deposition with its loss-of-function mutant displaying thicker interfascicular and xylary fiber cell walls but thinner vessel cell walls in inflorescence stems. To explore the exact function of KNAT7, class II KNOTTED1-LIKE HOMEOBOX (KNOX II) genes in Arabidopsis including KNAT3, KNAT4, and KNAT5 were studied together. By chimeric repressor technology, we found that both KNAT3 and KNAT7 repressors exhibited a similar dwarf phenotype. Both KNAT3 and KNAT7 genes were expressed in the inflorescence stems and the knat3 knat7 double mutant exhibited a dwarf phenotype similar to the repressor lines. A stem cross-section of knat3 knat7 displayed an enhanced irregular xylem phenotype as compared with the single mutants, and its cell wall thickness in xylem vessels and interfascicular fibers was significantly reduced. Analysis of cell wall chemical composition revealed that syringyl lignin was significantly decreased while guaiacyl lignin was increased in the knat3 knat7 double mutant. Coincidently, the knat3 knat7 transcriptome showed that most lignin pathway genes were activated, whereas the syringyl lignin-related gene Ferulate 5-Hydroxylase (F5H) was down-regulated. Protein interaction analysis revealed that KNAT3 and KNAT7 can form a heterodimer, and KNAT3, but not KNAT7, can interact with the key secondary cell wall formation transcription factors NST1/2, which suggests that the KNAT3-NST1/2 heterodimer complex regulates F5H to promote syringyl lignin synthesis. These results indicate that KNAT3 and KNAT7 synergistically work together to promote secondary cell wall biosynthesis., (© The Author(s) 2020. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Introduction: Youth with autism spectrum disorder (ASD) face high rates of unemployment, with unique challenges for military-dependent and -connected youth with ASD. This paper reports preliminary findings from Year One of a randomized waitlist controlled trial investigating the efficacy of the Project SEARCH + ASD Supports (PS + ASD) intervention model for military-dependent and -connected youth with ASD., Methods: Treatment group participants (n = 6) participated in internships at a military installation in the southeastern United States; waitlist group participants (n = 8) received special education transition services at their local high schools. Employment outcome data were collected at 12 months for both groups., Results: Fourteen unique internship experiences were developed across seven business partner organizations on the military installation during Year One. Five of six PS + ASD treatment group participants obtained competitive integrated employment for an overall employment rate of 83.3%. Four of the positions were federal jobs. None of the waitlist group participants obtained competitive integrated employment during the same period., Conclusions: Initial results are promising and suggest that the PS + ASD model may help to meet the transition needs of military-dependent and -connected youth with ASD and the employment needs of local military communities., (© Association of Military Surgeons of the United States 2019. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Cellulose synthase A genes (CesAs) are responsible for cellulose biosynthesis in plant cell walls. In this study, functions of secondary wall cellulose synthases PtrCesA4, PtrCesA7-A/B and PtrCesA8-A/B were characterized during wood formation in Populus trichocarpa (Torr. & Gray). CesA RNAi knockdown transgenic plants exhibited stunted growth, narrow leaves, early necrosis, reduced stature, collapsed vessels, thinner fiber cell walls and extended fiber lumen diameters. In the RNAi knockdown transgenics, stems exhibited reduced mechanical strength, with reduced modulus of rupture (MOR) and modulus of elasticity (MOE). The reduced mechanical strength may be due to thinner fiber cell walls. Vessels in the xylem of the transgenics were collapsed, indicating that water transport in xylem may be affected and thus causing early necrosis in leaves. A dramatic decrease in cellulose content was observed in the RNAi knockdown transgenics. Compared with wildtype, the cellulose content was significantly decreased in the PtrCesA4, PtrCesA7 and PtrCesA8 RNAi knockdown transgenics. As a result, lignin and xylem contents were proportionally increased. The wood composition changes were confirmed by solid-state NMR, two-dimensional solution-state NMR and sum-frequency-generation vibration (SFG) analyses. Both solid-state nuclear magnetic resonance (NMR) and SFG analyses demonstrated that knockdown of PtrCesAs did not affect cellulose crystallinity index. Our results provided the evidence for the involvement of PtrCesA4, PtrCesA7-A/B and PtrCesA8-A/B in secondary cell wall formation in wood and demonstrated the pleiotropic effects of their perturbations on wood formation., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Risk assessment have become essential in the prevention of cardiovascular disease. Even though risk prediction tools are recommended in the European guidelines, they are not adequately implemented in clinical practice. Risk prediction tools are meant to estimate prognosis in an unbiased and reliable way and to provide objective information on outcome probabilities. They support informed treatment decisions about the initiation or adjustment of preventive medication. Risk prediction tools facilitate risk communication to the patient and their family, and this may increase commitment and motivation to improve their health. Over the years many risk algorithms have been developed to predict 10-year cardiovascular mortality or lifetime risk in different populations, such as in healthy individuals, patients with established cardiovascular disease and patients with diabetes mellitus. Each risk algorithm has its own limitations, so different algorithms should be used in different patient populations. Risk algorithms are made available for use in clinical practice by means of - usually interactive and online available - tools. To help the clinician to choose the right tool for the right patient, a summary of available tools is provided. When choosing a tool, physicians should consider medical history, geographical region, clinical guidelines and additional risk measures among other things. Currently, the U-prevent.com website is the only risk prediction tool providing prediction algorithms for all patient categories, and its implementation in clinical practice is suggested/advised by the European Association of Preventive Cardiology.

A majority of the potential health benefits of green tea, including the potential to prevent cognitive decline, have been attributed to epigallocatechin gallate (EGCG). Sunrouge is a green tea cultivar that contains EGCG and several other bioactive components such as quercetin, myricetin, cyanidin and delphinidin. We compared the effects of Sunrouge and Yabukita, the most popular Japanese green tea cultivar, on cognitive function in the senescence-accelerated mouse Prone8. These mice were fed an experimental diet containing Sunrouge extract (SRE) or Yabukita extract (YBE). SRE feeding significantly prevented cognitive decline, whereas YBE feeding had little effect. Moreover, SRE feeding prevented elevation of the amyloid-β42 level while improving the gene expression of neprilysin and decreasing beta-site APP-cleaving enzyme 1 in the brain. These preventive effects of SRE against cognitive decline were attributed to the characteristic composition of Sunrouge and strongly suggest that consumption of this cultivar could protect against age-related cognitive decline.

High-speed counter-current chromatography (HSCCC) combined with macroporous resin (MR) column was successfully applied to the isolation and purification of four flavonoid glycosides from the medicinal herb Lotus plumule (LP). A polar two-phase solvent system composed of ethyl acetate-n-butanol-water (1:2:3, v/v/v) was selected by high-performance liquid chromatography (HPLC) and run on a preparative scale where the lower aqueous phase was used as the mobile phase with a head-to-tail elution mode. Quercetin-3-O-β-D-glucopyranoside (15 mg), isorhamnetin-3-O-β-D-glucopyranoside (13 mg), apigenin 6-C-β-D-glucopyranosyl-8-C-α-L-arabinopyranoside (18 mg) and apigenin 6,8-di-C-β-D-glucopyranoside (48 mg) were obtained in a one-step HSCCC separation from 240 mg of the sample. The purity of each compound was over 95% as determined by HPLC. Chemical structures of the isolated compounds were identified by electrospray ionization mass spectrometry (ESI-MS-MS) and nuclear magnetic resonance (NMR) methods. Moreover, the four compounds were isolated from LP for the first time., (© The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Aims: The aim of this study was to examine the antimicrobial properties of novel aqueous natural rapeseed oil/saline emulsions containing different soluble components of spruce resin., Methods and Results: The composition of aqueous resin emulsions was analysed by GC-MS and their antimicrobial properties were studied with challenge tests and with turbidometric assays. The emulsions were strongly antimicrobial against common Gram-positive and Gram-negative bacteria (including MRSA) as well as common yeasts. Furthermore, they inhibited the biofilm formation and eradicated the microbial biofilms on tested microbes. Characteristic for the emulsions was the presence of oxidized resin acids. Other main components present in emulsions, such as lignans and coumaric acids, were not antimicrobial, when tested separately., Conclusions: The results indicated that the oxidized resin acids were the antimicrobial components in the emulsions. Also, there appears to be a stoichiometric relationship between the number of resin acid molecules and the number microbe cells in the antimicrobial action., Significance and Impact of the Study: The fact that these solutions do not contain abietic acid, which is the main allergenic compound in resins, suggests that these solutions would be suitable, well-tolerated antimicrobials for various medical applications. The aqueous formulation will also allow the expansion of the use of these emulsions in from medical applications to the food preservatives and disinfectants., (© 2017 The Authors. Journal of Applied Microbiology published by John Wiley & Sons Ltd on behalf of the Society for Applied Microbiology.)

This research note considers how to track long-term trajectories of political discontent in Britain. Many accounts are confined to using either survey data drawn from recent decades or imperfect behavioral measures such as voting or party membership as indicators of political disengagement. We instead develop an approach that provides the long view on political disaffection. We first consider time-series data available from repeated survey measures. We next replicate historic survey questions to observe change in public opinion relative to earlier points in time. Finally, we use Stimson's (1991) dyad-ratios algorithm to construct an over-time index of political discontent that combines data from multiple poll series. This reveals rising levels of political discontent for both specific and diffuse measures of mass opinion. Our method and findings offer insights into the rising tide of disillusionment afflicting many contemporary democracies.

The response of Clostridium tyrobutyricum to butyric acid stress involves various stress-related genes, and therefore overexpression of stress-related genes can improve butyric acid tolerance and yield. Class I heat shock proteins (HSPs) play an important role in the process of protecting bacteria from sudden changes of extracellular stress by assisting protein folding correctly. The results of quantitative real-time PCR indicated that the Class I HSGs grpE, dnaK, dnaJ, groEL, groES, and htpG were significantly upregulated under butyric acid stress, especially the dnaK and groE operons. Overexpression of groESL and htpG could significantly improve the tolerance of C. tyrobutyricum to butyric acid, while overexpression of dnaK and dnaJ showed negative effects on butyric acid tolerance. Acid production was also significantly promoted by increased GroESL expression levels; the final butyric acid and acetic acid concentrations were 28.2 and 38% higher for C. tyrobutyricum ATCC 25755/groESL than for the wild-type strain. In addition, when fed-batch fermentation was carried out using cell immobilization in a fibrous-bed bioreactor, the butyric acid yield produced by C. tyrobutyricum ATCC 25755/groESL reached 52.2 g/L, much higher than that for the control. The improved butyric acid yield is probably attributable to the high GroES and GroEL levels, which can stabilize the biosynthetic machinery of C. tyrobutyricum under extracellular butyric acid stress.

Cross-national data production in social science research has increased dramatically in recent decades. Assessing the comparability of data is necessary before drawing substantive conclusions that are based on cross-national data. Researchers assessing data comparability typically use either quantitative methods such as multigroup confirmatory factor analysis or qualitative methods such as online probing. Because both methods have complementary strengths and weaknesses, this study applies both multigroup confirmatory factor analysis and online probing in a mixed-methods approach to assess the comparability of constructive patriotism and nationalism, two important concepts in the study of national identity. Previous measurement invariance tests failed to achieve scalar measurement invariance, which prohibits a cross-national comparison of latent means (Davidov 2009). The arrival of the 2013 ISSP Module on National Identity has encouraged a reassessment of both constructs and a push to understand why scalar invariance cannot be achieved. Using the example of constructive patriotism and nationalism, this study demonstrates how the combination of multigroup confirmatory factor analysis and online probing can uncover and explain issues related to cross-national comparability.

Scientists have been debating for centuries the nature of proper scientific methods. Currently, criticisms being thrown at data-intensive science are reinvigorating these debates. However, many of these criticisms represent long-standing conflicts over the role of hypothesis testing in science and not just a dispute about the amount of data used. Here, we show that an iterative account of scientific methods developed by historians and philosophers of science can help make sense of data-intensive scientific practices and suggest more effective ways to evaluate this research. We use case studies of Darwin's research on evolution by natural selection and modern-day research on macrosystems ecology to illustrate this account of scientific methods and the innovative approaches to scientific evaluation that it encourages. We point out recent changes in the spheres of science funding, publishing, and education that reflect this richer account of scientific practice, and we propose additional reforms.

We examined the cooperative effects of isoflavones and cello-oligosaccharides on daidzein metabolism and bone fragility in ovariectomized mice. Cello-oligosaccharides increased urinary equol and decreased O-desmethylangolensin. A combination of isoflavones and cello-oligosaccharides attenuated decreases in bone breaking force and stiffness caused by ovariectomy. Combination treatment with isofalvones and cello-oligosaccharides increases urinary equol/O-desmethylangolensin production ratio and prevents ovariectomy-induced abnormalities in bone strength.

Democratic responsiveness concerns the degree to which government policies match public preferences. Responsiveness studies typically use national surveys to characterize public opinion, but whether poll questions overlap with the policy agenda is unknown. The first of two empirical analyses presented here, with hundreds of issues on the national agenda in the United States from 1947 to 2000, reveals that public opinion is mostly unrelated to policy outcomes. The picture appears to be even more ominous-that is, opinion and policy are negatively related-on highly salient issues that attract media attention. A second study revisiting published work confirms that responsiveness patterns look different depending upon whether studies of opinion-policy connections (a) begin with survey data and then examine policy developments, or (b) begin with national legislative agenda issues and then examine survey data. Thus, conclusions about democratic responsiveness depend upon the issues that are examined, and often opinion surveys do not include questions about tangible public policy options. In that sense, future changes in democratic responsiveness might go undetected because scholars often lack data on what goes into the denominator of democracy.

This study examines the role of information in shaping public opinion in the context of support for education spending. While there is broad public support for increasing government funding for public schools, Americans tend to underestimate what is currently spent. We embed a series of experiments in a nationally representative survey administered in 2012 ( n = 2,993) to examine whether informing citizens about current levels of education spending alters public opinion about whether funding should increase. Providing information on per-pupil spending in a respondent's local school district reduces the probability that he or she will express support for increasing spending by 22 percentage points on average. Informing respondents about state-average teacher salaries similarly depresses support for salary increases. These effects are larger among respondents who underestimate per-pupil spending and teacher salaries by a greater amount, consistent with the idea that the observed changes in opinion are driven, at least in part, by informational effects, as opposed to priming alone.

Survey institutes recently have changed their measurement of generalized trust from the standard dichotomous scale to an 11-point scale. Additionally, numerous survey institutes use different question wordings: where most rely on the standard, fully balanced question (asking if "most people can be trusted or that you need to be very careful in dealing with people"), some use minimally balanced questions, asking only if it is "possible to trust people." By using two survey-embedded experiments, one with 12,009 self-selected respondents and the other with a probability sample of 2,947 respondents, this study evaluates the generalized trust question in terms of question wording and number of scale points used. Results show that, contrary to the more commonly used standard question format (used, for example, by the American National Election Studies and the General Social Survey), generalized trust is best measured with a minimally balanced question wording accompanied with either a seven- or an 11-point scale.

Aims: Currently, there is no direct method for detecting Clostridium thermocellum in the insoluble medium. In this study, a quantitative real-time PCR assay was developed for the direct growth detection of C. thermocellum at the single-cell level in lignocellulosic biomasses., Methods and Results: The assay targeted the cipA gene and was able to distinguish C. thermocellum from other species with good reproducibility which quantitative detection limit was 10 cell equivalents (CE) per reaction. OD600-based counting and qPCR quantification of C. thermocellum cultured in soluble medium were compared and an excellent consistency was revealed, indicating the appropriateness of the developed qPCR method. Analysis based on yellow affinity substrate and fermentation products may incorrectly estimate its population., Conclusions: The developed assay can serve as a specific, sensitive and reproducible method for the detection of C. thermocellum in lignocellulosic biomass at the single-cell level., Significance and Impact of the Study: With the ability to rapidly detect C. thermocellum, this method will contribute substantially to the understanding of the lignocellulosic biomass degradation mechanism. Moreover, it can also be applied to detect C. thermocellum growth in certain co-culture system for the understanding of the metabolic interactions., (© 2015 The Society for Applied Microbiology.)

Various economic and environmental sustainability concerns as well as consumer preference for bio-based products from natural sources have paved the way for the development and expansion of biorefining technologies. These involve the conversion of renewable biomass feedstock to fuels and chemicals using biological systems as alternatives to petroleum-based products. Filamentous fungi possess an expansive portfolio of products including the multifunctional organic acids itaconic, fumaric, and malic acids that have wide-ranging current applications and potentially addressable markets as platform chemicals. However, current bioprocessing technologies for the production of these compounds are mostly based on submerged fermentation, which necessitates physicochemical pretreatment and hydrolysis of lignocellulose biomass to soluble fermentable sugars in liquid media. This review will focus on current research work on fungal production of itaconic, fumaric, and malic acids and perspectives on the potential application of solid-state fungal cultivation techniques for the consolidated hydrolysis and organic acid fermentation of lignocellulosic biomass.

The class IV homeodomain leucine zipper transcription factor GLABRA2 (GL2) acts in a complex regulatory circuit that regulates the differentiation of trichomes in Arabidopsis thaliana. We describe a genetic interaction with HOMEODOMAIN GLABROUS11 (HDG11), previously identified as a negative regulator of trichome branching. gl2 hdg11 double mutants display enhanced trichome cell-type differentiation defects. Transgenic expression of HDG11 using the GL2 promoter partially suppresses gl2 trichome phenotypes. Vice versa, expression of GL2 under the control of its native promoter partially complements hdg11 ectopic branching. Since gl2 hdg11 and gl2 myb23 double mutants and the triple mutant display similar trichome differentiation defects, we investigated a connection to the R2R3-MYB transcription factor MYB23. We show that MYB23 transcript levels are significantly reduced in shoots from gl2 mutants and that GL2 can drive the expression of a MYB23-promoter fusion to green fluorescent protein. Yeast one-hybrid, chromatin immunoprecipitation, and in planta reporter gene experiments indicate that an L1-box in the MYB23 promoter acts as a GL2 binding site. Taken together, our findings reveal a functional redundancy between GL2 and HDG11, two homeodomain leucine zipper transcription factors previously thought to mediate opposing functions in trichome morphogenesis. A model is proposed in which GL2 transcript levels are maintained through a positive feedback loop involving GL2 activation of MYB23., (© 2014 American Society of Plant Biologists. All rights reserved.)

Background: Ependymoma is treated with maximal surgical resection and localized radiotherapy. Minimizing unnecessary exposure to radiation is of paramount importance for young children. Proton radiotherapy (PRT) spares healthy tissues outside the target region, but reports of clinical outcomes are scarce. We report outcomes for 70 patients treated with PRT for intracranial ependymoma., Methods: Seventy patients with localized ependymoma treated with involved-field PRT at the Massachusetts General Hospital between October 2000 and February 2011 were included., Results: Median age at diagnosis was 38 months (range, 3 mo-20 y). Nineteen (27%) patients had supratentorial ependymoma and 51(73%) had infratentorial ependymoma. Forty-six (66%) had gross total resection (GTR), and 24 (34%) had subtotal resection (STR). At a median follow-up of 46 months, 3-year local control, progression-free survival, and overall survival were 83%, 76%, and 95%, respectively. STR was significantly associated with worse progression-free survival (54% vs 88%, P = .001) and overall survival (90% vs 97% for GTR, P = .001). In a subset of patients (n = 14), mean intelligence was 108.5 at baseline and 111.3 after mean 2.05 years of follow-up. In a larger group of patients (n = 28), overall adaptive skills were 100.1 at baseline and 100.8 after 2.21 years of follow-up. Few patients developed evidence of growth hormone deficiency, hypothyroidism, or hearing loss., Conclusion: Outcomes for children treated with PRT compare favorably with the literature. STR correlated with inferior outcome. The young age at diagnosis and the proximity of critical structures in patients with ependymoma make PRT an ideal radiation modality.

This work comprises the structural characterization of Eucalyptus and Pinus radiata pulp fibers and their corresponding fibrillated materials, based on quantitative electron microscopy techniques. Compared to hardwood fibers, the softwood fibers have a relatively open structure of the fiber wall outer layers. The fibrillation of the fibers was performed mechanically and chemi-mechanically. In the chemi-mechanical process, the pulp fibers were subjected to a TEMPO-mediated oxidation to facilitate the homogenization. Films were made of the fibrillated materials to evaluate some structural properties. The thicknesses and roughnesses of the films were evaluated with standardized methods and with scanning electron microscopy (SEM), in backscattered electron imaging mode. Field-emission SEM (FE-SEM) and transmission electron microscopy (TEM) were performed to quantify the nanofibril morphology. In this study, we give additional and significant evidences about the suitability of electron microscopy techniques for quantification of nanofibril structures. In addition, we conclude that standard methods are not suitable for estimating the thickness of films having relatively rough surfaces. The results revealed significant differences with respect to the morphology of the fibrillated material. The differences are due to the starting raw material and to the procedure applied for the fibrillation.

Aims: To survey paperboard products from 17 US mills for bacterial populations and for bacteria potentially harmful to human health., Methods and Results: Culturable aerobic bacteria were isolated from paperboard products using selective and nonselective medium. Resulting colonies from samples from three regions of the United States were identified using fatty acid methyl ester analysis. Percentages of bacteria species found were Bacillus megaterium (47), Bacillus licheniformis (15), Bacillus pumilus (12), Paenibacillus macerans (5), Paenibacillus pabuli (3), Bacillus subtilis (2), Bacillus cereus (2), Bacillus coagulans (1), Bacillus circulans (1), Bacillus brevis (1), Bacillus thuringiensis (1), Paenibacillus polymyxa (1), Cellulomonas turbata (1), Cellulomonas flavigena (1), unidentified Bacillus sp. (3) and unidentified bacteria (1)., Conclusions: Recycled paperboard contained high populations of bacteria, and a positive correlation was found between recycle content and bacterial populations. Escherichia coli, Salmonella, Shigella or confirmed coliform bacteria were not found in any product., Significance and Impact of the Study: Populations of bacteria did not differ significantly from original counts over a 4-month period of dry storage, indicating that bacteria persist in paperboard over long periods and may re-enter the recycling process. The predominance of heat-tolerant endospore-forming bacteria explains the high bacteria counts found in paperboard made from recycled materials.

Aim: The purpose of this work was to study the feasibility of producing economic flame retardant bacterial cellulose (BC) and evaluating its behaviour in paper production., Methods and Results: This type of BC was prepared by Gluconacetobacter subsp. xylinus and substituting the glucose in the cultivation medium by glucose phosphate as a carbon source; as well as using corn steep liquor as a nitrogen source. The investigated processing technique did not dispose any toxic chemicals that pollute the surroundings or cause unacceptable effluents, making the process environmentally safe. The fire retardant behaviour of the investigated BC has been studied by non-isothermal thermogravimetric analysis (TGA & DTGA). The activation energy of each degradation stage and the order of degradation were estimated using the Coats-Redfern equation and the least square method. Strength, optical properties, and thermogravimetric analysis of BC-phosphate added paper sheets were also tested., Conclusions: The study confirmed that the use of glucose phosphate along with glucose was significant in the high yield production of phosphate containing bacterial cellulose (PCBC1); more so than the use of glucose phosphate alone (PCBC2). Incorporating 5% of the PCBC with wood pulp during paper sheet formation was found to significantly improve kaolin retention, strength, and fire resistance properties as compared to paper sheets produced from incorporating bacterial cellulose (BC)., Significance and Impact of the Study: This modified BC is a valuable product for the preparation of specialized paper, in addition to its function as a fillers aid.

We established a method for producing two types of xylooligosaccharides by treating kraft pulp derived from hardwood such as eucalyptus with xylanase, analyzed the structure by MALDI-TOF-MS/MS, and conducted an in vitro study on utilization by intestinal bacteria. One type was a neutral xylooligosaccharide (XOS), a xylooligomer in the D.P. range of dimer to approximately 17-mer, containing approximately 12% xylobiose. The other was an acidic xylooligosaccharide (U-XOS), a xylooligomer in the D.P. range of dimer to approximately 17-mer, with a major side chain of 4-O-methyl-glucuronic acid and other side chains of glucuronic acid and 2-O-galactopyranosyl-4-O-methyl-glucurono acid. The in vitro utilization study with intestinal bacteria showed that XOS was specifically utilized by Bifidobacterium, whereas U-XOS was utilized by only a few types of bacteria. In a fecal batch culture, both XOS and U-XOS selectively increased the population of Bifidobacterium. The present study proposes a new method for industrial production of XOS and U-XOS, and indicates the utility and functionality.

A major barrier to the commercialization of somatic embryogenesis technology in loblolly pine (LP, Pinus taeda L.) is recalcitrance of some high-value crosses to initiate embryogenic tissue and to continue early-stage somatic embryo growth. Developing initiation and multiplication media that resemble the seed environment may decrease this recalcitrance. Sugar and sugar alcohol analyses were performed weekly throughout the sequence of seed development for female gametophyte and zygotic embryo tissues to determine physiologic concentrations (Pullman, G.S. and M. Buchanan. 2008. Identification and quantitative analysis of stage-specific carbohydrates in LP (Pinus taeda) zygotic embryo and female gametophyte tissues. Tree Physiol. 28:985-996). Major differences in stage-specific sugars were observed. A simple bioassay was used to evaluate the potential growth promotion of individual carbohydrates added to initiation or multiplication media at physiologic concentrations. Seventeen sugars were screened. Compounds showing statistically significant increases in early-stage embryo growth were then tested for the ability to increase the initiation of LP. d-xylose and d-chiro-inositol produced statistically significant increases in early-stage embryo growth. When tested for improved initiation in P. taeda, Pseudotsuga menziesii (mirb) Franco and Picea abies L., Karst., d-xylose increased the averages of initiation by 6.5%, 7.3% and 16.7%, respectively. d-chiro-inositol increased the initiation in P. taeda by 7.3% in one test but not in the other, whereas in P. menziesii the initiation increases averaged 8.4% in two tests. Analyses of sugars and sugar alcohols in the seed environment coupled with a bioassay to screen potential media supplements for protocol improvement resulted in statistically significant increases in embryogenic tissue initiation for several coniferous species.

Stage-specific analyses of starch and 18 sugars, including pentoses, hexoses, disaccharides, trisaccharides, oligosaccharides and sugar alcohols, were made throughout seed development for zygotic embryo and female gametophyte (FG) tissues of loblolly pine (Pinus taeda L.). Tissue was most often analyzed in triplicate from two open-pollinated families grown in different locations and sampled in different years. Carbohydrates were analyzed by enzymatic assay, high performance liquid chromatography or gas chromatography/mass spectrometry. For all carbohydrates quantified, peak concentrations were higher in embryo tissue than in FG tissue. Significant changes in starch and sugar concentrations occurred over time, with both seed collections showing similar trends in temporal changes. Although concentrations were not always similar, embryo and FG tissues generally showed similar patterns of change in starch and sugar concentrations over time. Total starch concentration was highest during early seed development and decreased as development progressed. The major sugars contributing to osmotic potential during early seed development were D-pinitol, sucrose, fructose and glucose. During mid-seed development, D-pinitol, sucrose, fructose, glucose, melibiose and raffinose provided major contributions to the osmotic environment. During late seed development, sucrose, raffinose, melibiose, stachyose and fructose were the major contributors to osmotic potential. These data suggest stage-specific media composition for each step in the somatic embryogenesis protocol.

Recently we demonstrated that in inflammatory bowel disease (IBD) macrophage-oxidative burst activity is increased and NADPH oxidase mRNA is induced. The herbal phenylethanoid acteoside isolated from Plantago lanceolata L. was shown to exhibit anti-oxidative potential. Using the dextran sulphate sodium (DSS)-induced colitis model, in this study we have assessed whether systemic application of acteoside affects colitis. Colitis was induced by DSS in Balb/c mice. Treatment with acteoside (120, 600 microg/mouse/day) was performed intraperitoneally. The colon lengths were determined. Colonic tissue was scored histologically (max. score 8) by a blinded investigator. T cells isolated from mesenteric lymph nodes (MLN) were stimulated with anti-CD3 antibody in the presence of interleukin (IL)-2 (final concentration 10 U/ml). After incubation for 24 h, IL-1beta, IL-6, IL-12 tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma levels in supernatants were analysed by the beadlyte cytokine detection system. Histological scoring of colonic tissue revealed that application of acteoside was followed by a significantly improved histological score. In acute colitis the histological score was 3.2 with acteoside versus 5.2 with phosphate-buffered saline (PBS) (P < 0.02). In chronic colitis both 120 microg (3.3 versus 5.2) or 600 microg acteoside (3.0 versus 5.2) significantly ameliorated colitis (both P < 0.02). Stimulated MLN from mice with chronic DSS-induced colitis treated with acteoside showed a significant down-regulation of IFN-gamma secretion (195 pg/ml with 600 microg acteoside versus 612 pg/ml with PBS, P < 0.02). Inhibition of oxidative burst activity with acteoside reduced mucosal tissue damage in DSS colitis and could be a therapeutic alternative for IBD treatment. Further studies of this agent are warranted.

Juvenile wood (JW) of conifers is often associated with compression wood (CW), with which it is sometimes believed to be identical. To determine whether JW and CW can be distinguished metabolically, we compared gas chromatographic profiles of 25 polar metabolites from rooted cuttings of a single loblolly pine (Pinus taeda L.) clone raised in controlled environment chambers and subject to three treatments: (1) grown erect with minimal wind sway (control); (2) swayed by wind from oscillating fans; and (3) with 30-cm growth increments successively bent at an angle of 45 degrees to the vertical. Profiles were compared by principal component analysis. Substantial increases in abundances of coniferin and p-glucocoumaryl alcohol separated immature JW-forming xylem tissues of the control trees from the CW-forming xylem of the bent and swayed trees.

Transgenic Arabidopsis thaliana plants carrying a single copy of integrated DNA can be identified by single-step genomic polymerase chain reaction. The reaction employs two sets of primer pairs with the same melting temperature that amplify the amplicons derived from the integrated T-DNA together with those from an endogenous single-copy gene as a reference. When the band intensity ratio is one, this means that the transgenic plants are carrying a single copy of the integrated gene per haploid.

In order to save energy during the pulp making process, we tried to use white-rot basidiomycete, Trametes hirsuta, which degrades lignin efficiently. But a decrease in paper strength caused by cellulolytic activity ruled this out for practical application. Since the cellulolytic activity of the fungus must be decreased, we purified and characterized a cellobiose dehydrogenase (CDH) that was reported to damage pulp fiber. The CDH in the culture filtrate of C. hirsutus was purified by freeze-thawing and chromatographic methods. The pI of the enzyme was 4.2 and its molecular weight was 92 kDa. The optimal temperature and pH of the enzyme were 60-70 degrees C and 5.0 respectively. Since the purified CDH decreased the viscosity of pulp in the presence of Fe(III) and cellobiose, it was shown that the suppression of CDH should be an effective way to reduce cellulose damage.

Acrylamide has a range of toxicological hazards including neurotoxicity and reproductive toxicity; however, occupational risk management is driven by its genotoxic and carcinogenic potential (it is classified within the EU as a Category 2 carcinogen, R45 and Category 2 mutagen, R46). Since there is the potential for skin absorption and systemic toxicity, biological monitoring may be a useful aid for the assessment of exposure via inhalation, ingestion and dermal absorption. However, there are currently no biological monitoring guidance values (BMGVs). This study describes an extensive survey of potential workplace exposure to acrylamide at the Ciba (Bradford) site to gather data suitable for a BMGV. This manufacturing site is typical within the industry as a whole and includes a cross section of activities and tasks representative of acrylamide exposure. Acrylamide is used in the manufacture of polyacrylamide based products for applications in water treatment; oil and mineral extraction; paper, paint and textile processes. Workers (62 plus 6 controls) with varying potential exposures provided a total of 275 pre shift and 247 post-shift urine samples along with 260 personal air samples. A small non-exposed control group was similarly monitored. Urine samples were analysed for S-carboxyethyl-cysteine (CEC). Airborne, surface and glove samples were analysed for acrylamide. Inhalation exposures were well controlled with values consistently below one-tenth of the UK Workplace Exposure Limit. Engineering controls, personal protective equipment and work practice, all contributed to good control of occupational exposure. CEC was found in urine samples from both exposed workers and non-occupationally exposed controls. At the low levels of exposure found, smoking made a significant contribution to urinary CEC levels. Nevertheless a correlation between urinary CEC and airborne acrylamide was found. A mixed effects model incorporating inhalation concentrations of acrylamide and smoking habits could predict some of the variation in observed post-shift urine results but could be improved through the use of additional surface contamination data. However, the data does not suggest that dermal absorption was a major contributor to the systemic dose. Based on the 90th percentile of the data, inclusive of the effects of smoking and environmental factors, a value of 4 mmol mol(-1) creatinine is proposed as a pragmatic BMGV associated with good occupational hygiene practice and control of workplace exposure. CEC in urine analysis has the utility for routine use as a means to estimate biological uptake where there is a potential for significant exposure or loss of workplace control.

Hydrophilic knotwood extracts from 18 wood species were assessed in disc diffusion and liquid culture tests for antibacterial effects against three species of paper mill bacteria. The Pinus sylvestris, P. resinosa, P. contorta, and P. banksiana extracts decreased or inhibited bacterial growth. The susceptibility order was P. sylvestris > P. resinosa > P. contorta > P. banksiana, correlating with the concentrations of pinosylvin and pinosylvin monomethyl ether in these wood species. Also, Pseudotsuga menziesii and Thuja occidentalis extracts had a small inhibitory effect. The Gram-positive Bacillus coagulans was more susceptible to the extracts than the Gram-negative Burkholderia multivorans and Alcaligenes xylosoxydans. The main components in the Pinus knotwood extracts were pinosylvin monomethyl ether and pinosylvin, suggesting these to be the active components. Therefore, pure pinosylvin, pinosylvin monomethyl ether, and dihydro-pinosylvin monomethyl ether were also tested. All compounds showed antibacterial effects. However, higher concentrations were needed for these pure compounds than for the knotwood extracts. Pinosylvin had stronger antibacterial effects than pinosylvin monomethyl ether. This work shows that knotwood extracts, especially from Pinus species, have a potential for use as natural biocides in papermaking.

Aims: We investigated the antimicrobial activities of eucalyptus leaf extracts to find effective antibacterial agents., Methods and Results: The antimicrobial activities of leaf extracts from 26 species of eucalyptus were measured. Extracts of Eucalyptus globulus, E. maculata and E. viminalis significantly inhibited the growth of six Gram-positive bacteria (Staphylococcus aureus, MRSA, Bacillus cereus, Enterococcus faecalis, Alicyclobacillus acidoterrestris, Propionibacterium acnes), and of a fungus (Trichophyton mentagrophytes), but they did not show strong antibacterial activity against Gram-negative bacteria (Escherichia coli, Pseudomonas putida). 2',6'-dihydroxy-3'-methyl-4'-methoxy-dihydrochalcone, eucalyptin and 8-desmethyl-eucalyptin, isolated from E. maculata extracts, exhibited potent antimicrobial activities against seven micro-organisms with minimum inhibitory concentrations (MIC) ranging from 1.0 to 31 mg l(-1)., Conclusions: The eucalyptus extracts and three compounds from E. maculata were found to be effective against micro-organisms that cause food poisoning, acne and athlete's foot., Significance and Impact of the Study: This study shows potential uses of extracts from E. globulus, E. maculata and E. viminalis, and antimicrobial compounds isolated from E. maculata.

An efficient transformation system for the basidiomycete Coriolus hirsutus was developed. A double-auxotrophic mutant of C. hirsutus, deficient both in ornithine carbamoyltransferase (OCTase) and 3-isopropylmalate dehydrogenase (3-IPM dehydrogenase), was transformed to Arg+ with each allelic type of the C. hirsutus genomic OCTase gene (arg1) newly cloned. The transformation frequency of 10(3)-10(4) transformants per mug DNA per 10(6)-10(7) oidial protoplasts was reached. Southern blots showed that the transforming DNA was integrated into chromosomal DNA with multi-copies. The Arg+ phenotype of the transformants was stably inherited through mitosis.

Lignin is one of the most abundant biopolymers, and it has a complex racemic structure. It may be formed by a radical polymerization initiated by redox enzymes, but much remains unknown about the process, such as how molecules as large as enzymes can generate the compact structure of the lignified plant cell wall. We have synthesized lignin oligomers according to a new concept, in which peroxidase is never in direct contact with the lignin monomers coniferaldehyde and coniferyl alcohol. Instead, manganese oxalate worked as a diffusible redox shuttle, first being oxidized from Mn(II) to Mn(III) by a peroxidase and then being reduced to Mn(II) by a simultaneous oxidation of the lignin monomers to radicals that formed covalent linkages of the lignin type. Furthermore, a high molecular mass polymer was generated by oxidation of coniferyl alcohol by Mn(III) acetate in a dioxane and water mixture. This polymer was very similar to natural spruce wood lignin, according to its NMR spectrum. The possible involvement of a redox shuttle/peroxidase system in lignin biosynthesis is discussed.

Abietane terpenoid-degrading organisms include Sphingomonas spp which inhabit natural environments and biological treatment systems. An isolate from the high Arctic indicates that these organisms occur far from trees which synthesize abietanes and suggests that some of these organisms can occupy a niche in hydrocarbon-degrading soil communities. Abietane-degrading Sphingomonas spp provide additional evidence that the phylogeny of this genus is independent of the catabolic capabilities of its members. Studies of Sphingomonas sp DhA-33 demonstrate that biological treatment systems for pulp mill effluents have the potential to mineralize abietane resin acids. On the other hand, these studies indicate that some chlorinated dehydroabietic acids are quite recalcitrant. Strain DhA-33 grows relatively well on some chlorinated dehydroabietic acids but transforms others to stable metabolites. Using strain DhA-33, a novel method was developed to measure the metabolic activity of an individual population within a complex microbial community. Oligonucleotide hybridization probes were used to assay the 16S rRNA:rDNA ratio of DhA-33 as it grew in an activated sludge community. However, this method proved not to be sufficiently sensitive to measure naturally occurring resin acid-degrading populations. We propose that the same approach can be modified to use more sensitive assays.

Deuterium- and tritium-labeled gibberellins (GAs) were applied to stems of 3-year-old Eucalyptus globulus Labill. saplings and 9-month-old potted seedlings. Cambial region tissues surrounding the application point were collected 6, 24 or 48 h later. Twenty-four hours after application of 5 &mgr;g of [(2)H(2)]GA(20), 7% of the cambial region GA(20) pool, 7% of the GA(1) pool and 58% of the GA(29) pool were labeled with deuterium based on selected ion monitoring of purified extracts subjected to gas chromatography-mass spectrometry. The relatively low percent dilution of endogenous GAs by [(2)H(2)]GAs suggests that the exogenous application of [(2)H(2)]GA(20) did not result in substrate overloading, indicating that these conversions probably occur naturally within cambial region tissues. Extracts from similar cambial region tissues fed tritium-labeled GAs were sequentially fractionated by SiO(2) partition chromatography, C(18) reversed phase HPLC and N(CH(3))(2) HPLC. The radioactivity profiles indicated metabolism of GA(20) to GA(1) and GA(29), GA(1) conversion to GA(8), GA(4) to GA(34) and GA(9) to GA(51). Gibberellins GA(34), GA(51) and GA(29) are C-2beta-hydroxylated catabolites of low biological activity, whereas GA(1) and GA(4) are probably effectors of growth in the Eucalyptus stem and shoot. Evidence for C-13 hydroxylation of GA(4) to GA(1), GA(9) to GA(4) or GA(9) to GA(20) in the stem was inconclusive. Thus, although GA(4) and GA(9) are native to cambial region tissues, GA(1) is probably not produced from them in significant quantities. We conclude that the early C-13-hydroxylation pathway; i.e., conversion of GA(19) to GA(20) to GA(1), is the major pathway of GA(1) biosynthesis.

In Britain, the law places duties on employers and suppliers to provide information to ensure health and safety of employees, so far as is reasonably practicable, and there are regulations for the appointment of employees' safety representatives which employers are required to consult. A recent survey by HSE has shown that suppliers are the most important source of information on substances. However, the flow of information is often obstructed by barriers between the suppliers and the companies, and within organisations. Safety representatives, where they exist, are often better informed than employers, and in companies with safety representatives accident rates are lower. Information from suppliers can be inappropriate for the end use, and the goal-setting approach which has dominated in recent years may not help the non-expert employer. We welcome HSE's move to more specific control guidance for chemicals.

The extent of genetic identity observed in the young individuals which were micropropagated from a single Eucalyptus individual was analyzed by using DNA-fingerprinting. Among 40,000 tissue-cultured-seedings of E.citriodora, 200 plants were randomly chosen so that each total DNA might be extracted from their leaves. Using these DNAs as template, PCR was performed with some primers we found in advance that leads polymorphism for DNA of E. citriodora. In this study, all over the 200 cases, the band pattern formed cDNA fragment on a gel after electrophoresis was the identical one mutually.

We found a novel PCR-primer which can be used for the identification on "elite-tree-selection". This primer was designed for selective hybridization at the both ends of microsatellite loci, which is well known as one of the most highpervariable region of DNA. After PCR-fingerprinting on five Eucalyptus species (E. globulus, E.citriodora, E.grandis, E. maidenii, E.bicostata), with our primer, DNA-polymorphism was observed all over the cases.

Ethanol production from spent sulphite pulping liquor (SSL) was compared for four different yeasts. A second strain of S. cerevisiae as well as a 2-deoxyglucose-resistant strain formed through protoplast fusions between S. uvarum and S. diastaticus produced up to 27% more ethanol from SSL fortified with hydrolysis sugars than was produced by S. cerevisiae. The incremental improvement in ethanol yield appeared to vary with the degree of fortification, ranging from 5.8% for unfortified SSL, to 27% for the highest level of fortification tested. Decreasing fermentation rates were observed for SSL fortified with glucose, mannose and galactose, respectively. Sugar uptake rates in SSL fortified with glucose, galactose and mannose were 6.8, 2.8 and 2.0 g L-1 h-1, respectively. However, when these sugars were fermented along with a glucose cosubstrate, the rate at which the combined glucose/mannose medium was fermented was nearly identical to that of the glucose control.

A total DNA extracted from Eucalyptus globlus, E. citriodora, or a subspecies of E. gunnii (Maruha Yu-kari) being used as an template, Polymerase Chain Reaction (PCR) was performed with 40 kinds of commercially available random primers (Operon, OPA-01-20 and OPB-01-20). After electrophoresis of PCR-products on agarose gel, we analyzed the band pattern formed by DNA-fragments with 0.5-3.0 kbp as molecular weight. As a remuneration for the examination in which forty 10-mer arbitrary oligonucleotides were tried to be used as primers in PCR for each species, we found that 9 primers (for E. globlus) leads polymorphism, 6 primers for E. citriodora, and 6 primers for Maruha Yu-kari.

The introduction of chimeric sense and antisense gene constructs for 4-coumarate:coenzyme A ligase into tobacco plants caused the reduction of the 4CL activity in the transgenic plants. In the transgenic plants, the cell walls of the xylem tissue in stems were brown and the molecular structure of lignin in the colored cell walls was dramatically different from that in the control plants. Analysis with different types of stain revealed that levels of cinnamyl aldehyde residues and syringyl units in lignin were depressed in the brownish cell walls. Furthermore, the lignin content in colored tissue was lower than that in the normal tissue. Our results indicate that 4CL has important roles in the determination of the composition and the amount of lignin in tobacco plants.