Arginine Decarboxylase from Lathyrus sativus Seedlings Purification and Properties

Arginine decarboxylase which makes its appearance in Lathyrus sativus seedlings after 24 h of seed germination reaches its highest level around 5–7 days, the cotyledons containing about 60% of the total activity in the seedlings at day 5. The cytosol enzyme was purified 977-fold from whole seedlings by steps involving manganese chloride treatment, ammonium sulphate and acetone fractionations, positive adsorption on alumina C-γ gel, DEAE-Sephadex chromatography followed by preparative disc gel electrophoresis. The enzyme was shown to be homogeneous by electrophoretic and immunological criteria, had a molecular weight of 220000 and appears to be a hexamer with identical subunits. The optimal pH and temperature for the enzyme activity were 8.5 and 45 °C respectively. The enzyme follows typical Michaelis-Menten kinetics with a Km value of 1.73 mM for arginine. Though Mn2+ at lower concentrations stimulated the enzyme activity, there was no dependence of the enzyme on any metal for the activity. The arginine decarboxylase of L. sativus is a sulfhydryl enzyme. The data on co-factor requirement, inhibition by carbonyl reagents, reducing agents and pyridoxal phosphate inhibitors, and a partial reversal by pyridoxal phosphate of inhibition by pyridoxal · HCl suggests that pyridoxal 5'-phosphate is involved as a co-factor for the enzyme. The enzyme activity was inhibited competitively by various amines including the product agmatine. Highest inhibition was obtained with spermine and arcain. The substrate analogue, l-canavanine, homologue l-homoarginine and other basic amino acids like l-lysine and l-ornithine inhibited the enzyme activity competitively, homoarginine being the most effective in this respect.

Vanadate stimulates oxidation of NADH to generate hydrogen peroxide

Vanadate in the polymeric form of decavanadate, but not other forms, stimulated oxidation of NADH to NAD+ NADPH was also oxidized with comparable rates. This oxidation of NADH was accompanied by uptake of oxygen and generated hydrogen peroxide with the following stoichiometry: NADH + H+ + O2 → NAD+ + H2O2. The reaction followed second-order kinetics. The rate was dependent on the concentration of both NADH and vanadate and increased with decreasing pH. The reaction had an obligatory requirement for phosphate ions. Esr studies in the presence of the spin trap dimethyl pyrroline N oxide indicated the involvement of Superoxide anion as an intermediate. The reaction was sensitive to Superoxide dismutase and other scavengers of superoxide anions.

Agronomic benefits and risks associated with the irrigated peanut–maize production system under a changing climate in northern Australia

With the aim of increasing peanut production in Australia, the Australian peanut industry has recently considered growing peanuts in rotation with maize at Katherine in the Northern Territory—a location with a semi-arid tropical climate and surplus irrigation capacity. We used the well-validated APSIM model to examine potential agronomic benefits and long-term risks of this strategy under the current and warmer climates of the new region. Yield of the two crops, irrigation requirement, total soil organic carbon (SOC), nitrogen (N) losses and greenhouse gas (GHG) emissions were simulated. Sixteen climate stressors were used; these were generated by using global climate models ECHAM5, GFDL2.1, GFDL2.0 and MRIGCM232 with a median sensitivity under two Special Report of Emissions Scenarios over the 2030 and 2050 timeframes plus current climate (baseline) for Katherine. Effects were compared at three levels of irrigation and three levels of N fertiliser applied to maize grown in rotations of wet-season peanut and dry-season maize (WPDM), and wet-season maize and dry-season peanut (WMDP). The climate stressors projected average temperature increases of 1°C to 2.8°C in the dry (baseline 24.4°C) and wet (baseline 29.5°C) seasons for the 2030 and 2050 timeframes, respectively. Increased temperature caused a reduction in yield of both crops in both rotations. However, the overall yield advantage of WPDM increased from 41% to up to 53% compared with the industry-preferred sequence of WMDP under the worst climate projection. Increased temperature increased the irrigation requirement by up to 11% in WPDM, but caused a smaller reduction in total SOC accumulation and smaller increases in N losses and GHG emission compared with WMDP. We conclude that although increased temperature will reduce productivity and total SOC accumulation, and increase N losses and GHG emissions in Katherine or similar northern Australian environments, the WPDM sequence should be preferable over the industry-preferred sequence because of its overall yield and sustainability advantages in warmer climates. Any limitations of irrigation resulting from climate change could, however, limit these advantages.

Silica Vesicle Nanovaccine Formulations Stimulate Long-Term Immune Responses to the Bovine Viral Diarrhoea Virus E2 Protein

Bovine Viral Diarrhoea Virus (BVDV) is one of the most serious pathogen, which causes tremendous economic loss to the cattle industry worldwide, meriting the development of improved subunit vaccines. Structural glycoprotein E2 is reported to be a major immunogenic determinant of BVDV virion. We have developed a novel hollow silica vesicles (SV) based platform to administer BVDV-1 Escherichia coli-expressed optimised E2 (oE2) antigen as a nanovaccine formulation. The SV-140 vesicles (diameter 50 nm, wall thickness 6 nm, perforated by pores of entrance size 16 nm and total pore volume of 0.934 cm(3)g(-1)) have proven to be ideal candidates to load oE2 antigen and generate immune response. The current study for the first time demonstrates the ability of freeze-dried (FD) as well as non-FD oE2/SV140 nanovaccine formulation to induce long-term balanced antibody and cell mediated memory responses for at least 6 months with a shortened dosing regimen of two doses in small animal model. The in vivo ability of oE2 (100 mu g)/SV-140 (500 mu g) and FD oE2 (100 mu g)/SV-140 (500 mu g) to induce long-term immunity was compared to immunisation with oE2 (100 mu g) together with the conventional adjuvant Quil-A from the Quillaja saponira (10 mu g) in mice. The oE2/SV-140 as well as the FD oE2/SV-140 nanovaccine generated oE2-specific antibody and cell mediated responses for up to six months post the final second immunisation. Significantly, the cell-mediated responses were consistently high in mice immunised with oE2/SV-140 (1,500 SFU/million cells) at the six-month time point. Histopathology studies showed no morphological changes at the site of injection or in the different organs harvested from the mice immunised with 500 mu g SV-140 nanovaccine compared to the unimmunised control. The platform has the potential for developing single dose vaccines without the requirement of cold chain storage for veterinary and human applications.

Use of phosphorus fertiliser for increased productivity of legume-based sown pastures in the Brigalow Belt region – a review

The Brigalow Belt bioregion of southern and central Queensland supports a large percentage of northern Australia's sown pastures and beef herd. The Brigalow soils were widely thought to have adequate phosphorus (P) for cropping, sown pastures and grazing animals, which has led to almost no use of P fertiliser on sown pastures. The majority of pastures established in the region were sown with tropical grasses only (i.e. no legumes were sown). Under grass-only pastures, nitrogen (N) mineralisation rates decline with time since establishment as N is 'tied-up' in soil organic matter. This process leads to a significant decline in pasture and animal productivity and is commonly called 'pasture rundown'. Incorporating pasture legumes has been identified as the best long-term solution to improve the productivity of rundown sown grass pastures. Pasture legumes require adequate P to grow well and fix large amounts of N to increase the productivity of rundown sown grass pastures. Producers and farm advisors have traditionally thought that P fertiliser is not cost-effective for legume-based improved pastures growing on inland areas of Queensland despite there being little, if any, data on production responses or their economic outcomes. Recent studies show large and increasing areas of low plant available soil P and large responses by pasture legumes to P fertiliser on Brigalow soils. The economic analysis in this scoping study indicates potential returns of 9–15% on extra funds invested from the application of P fertiliser, when establishing legumes into grass pastures on low P soils (i.e. lower than the critical P requirement of the legume grown). Higher returns of 12–24% may be possible when adding P fertiliser to already established grass/legume pastures on such soils. As these results suggest potential for significant returns from applying P fertiliser on legume pastures, it is recommended that research be conducted to better quantify the impacts of P fertiliser on productivity and profit. Research priorities include: quantifying the animal production and economic impact of fertilising legume-based pastures in the sub-tropics for currently used legumes; quantifying the comparative P requirements and responses of available legume varieties; understanding clay soil responses to applied P fertiliser; testing the P status of herds grazing in the Brigalow Belt; and quantifying the extent of other nutrient deficiencies (e.g. sulphur and potassium) for legume based pastures. Development and extension activities are required to demonstrate the commercial impacts of applying P fertiliser to legume based pastures.

Electronic Building Passport - Final Report: Project 2 National Energy Efficient Building Project Phase 2

The National Energy Efficient Building Project (NEEBP) Phase One report, published in December 2014, investigated “process issues and systemic failures” in the administration of the energy performance requirements in the National Construction Code. It found that most stakeholders believed that under-compliance with these requirements is widespread across Australia, with similar issues being reported in all states and territories. The report found that many different factors were contributing to this outcome and, as a result, many recommendations were offered that together would be expected to remedy the systemic issues reported. To follow up on this Phase 1 report, three additional projects were commissioned as part of Phase 2 of the overall NEEBP project. This Report deals with the development and piloting of an Electronic Building Passport (EBP) tool – a project undertaken jointly by pitt&sherry and a team at the Queensland University of Technology (QUT) led by Dr Wendy Miller. The other Phase 2 projects cover audits of Class 1 buildings and issues relating to building alterations and additions. The passport concept aims to provide all stakeholders with (controlled) access to the key documentation and information that they need to verify the energy performance of buildings. This trial project deals with residential buildings but in principle could apply to any building type. Nine councils were recruited to help develop and test a pilot electronic building passport tool. The participation of these councils – across all states – enabled an assessment of the extent to which these councils are currently utilising documentation; to track the compliance of residential buildings with the energy performance requirements in the National Construction Code (NCC). Overall we found that none of the participating councils are currently compiling all of the energy performance-related documentation that would demonstrate code compliance. The key reasons for this include: a major lack of clarity on precisely what documentation should be collected; cost and budget pressures; low public/stakeholder demand for the documentation; and a pragmatic judgement that non-compliance with any regulated documentation requirements represents a relatively low risk for them. Some councils reported producing documentation, such as certificates of final completion, only on demand, for example. Only three of the nine council participants reported regularly conducting compliance assessments or audits utilising this documentation and/or inspections. Overall we formed the view that documentation and information tracking processes operating within the building standards and compliance system are not working to assure compliance with the Code’s energy performance requirements. In other words the Code, and its implementation under state and territory regulatory processes, is falling short as a ‘quality assurance’ system for consumers. As a result it is likely that the new housing stock is under-performing relative to policy expectations, consuming unnecessary amounts of energy, imposing unnecessarily high energy bills on occupants, and generating unnecessary greenhouse gas emissions. At the same time, Councils noted that the demand for documentation relating to building energy performance was low. All the participant councils in the EBP pilot agreed that documentation and information processes need to work more effectively if the potential regulatory and market drivers towards energy efficient homes are to be harnessed. These findings are fully consistent with the Phase 1 NEEBP report. It was also agreed that an EBP system could potentially play an important role in improving documentation and information processes. However, only one of the participant councils indicated that they might adopt such a system on a voluntary basis. The majority felt that such a system would only be taken up if it were: - A nationally agreed system, imposed as a mandatory requirement under state or national regulation; - Capable of being used by multiple parties including councils, private certifiers, building regulators, builders and energy assessors in particular; and - Fully integrated into their existing document management systems, or at least seamlessly compatible rather than a separate, unlinked tool. Further, we note that the value of an EBP in capturing statistical information relating to the energy performance of buildings would be much greater if an EBP were adopted on a nationally consistent basis. Councils were clear that a key impediment to the take up of an EBP system is that they are facing very considerable budget and staffing challenges. They report that they are often unable to meet all community demands from the resources available to them. Therefore they are unlikely to provide resources to support the roll out of an EBP system on a voluntary basis. Overall, we conclude from this pilot that the public good would be well served if the Australian, state and territory governments continued to develop and implement an Electronic Building Passport system in a cost-efficient and effective manner. This development should occur with detailed input from building regulators, the Australian Building Codes Board (ABCB), councils and private certifiers in the first instance. This report provides a suite of recommendations (Section 7.2) designed to advance the development and guide the implementation of a national EBP system.

Halvennettu työ : Pätkätyö ja sukupuoli sopimusyhteiskuntaa edeltävissä työmarkkinakäytännöissä

Unvalued Work. Gender and fragmented labour before national collective bargaining Systematically irregular work creates economic and social insecurity. A novelty? To think that globalisation results in unprecedented labour conditions turns out to be questionable when the idea is put in perspective. In the light of history there is nothing new in the frequency of today s short-term employment, for instance, ranking genders in labour relations in an old custom. Unvalued Work (Halvennettu työ) examines the regulation and management of labour before the time of collective bargaining. In the study present trends engage in a dialogue with empirical findings from the past. Preventing trade unions to take the initiative has been and remains an employer interest. The analysis focuses on female employment in the 1920s and 1930s. The inferences challenge to ask on what conditions the history of Finnish labour relations warrants on the whole speaking of contractual security, stable earnings and regular waged work that provides livelihood. Success in selling one s labour is not synonymous with good employment that yields decent income. Juxtaposing labour relations between the world wars and the 21st century helps us to understand the currently transforming labour market. Present policies are informed by past choices and patterns of thought. Unvalued Work (Halvennettu työ) offers instruments for making sense of today s labour relations.

Normalized Two Stage SVQ for Minimum Complexity Wide-band LSF Quantization

We develop a two stage split vector quantization method with optimum bit allocation, for achieving minimum computational complexity. This also results in much lower memory requirement than the recently proposed switched split vector quantization method. To improve the rate-distortion performance further, a region specific normalization is introduced, which results in 1 bit/vector improvement over the typical two stage split vector quantizer, for wide-band LSF quantization.

Essays on the Economics of Intellectual Property Rights

Väitöskirjassani tarkastelen informaatiohyödykkeiden ja tekijänoikeuksien taloustiedettä kahdesta eri perspektiivistä. Niistä ensimmäinen kuuluu endogeenisen kasvuteorian alaan. Väitöskirjassani yleistän ”pool of knowledge” -tyyppisen endogeenisen kasvumallin tilanteeseen, jossa patentoitavissa olevalla innovaatiolla on minimikoko, ja jossa uudenlaisen tuotteen patentoinut yritys voi menettää monopolinsa tuotteeseen jäljittelyn johdosta. Mallin kontekstissa voidaan analysoida jäljittelyn ja innovaatioilta vaaditun ”minimikoon” vaikutuksia hyvinvointiin ja talouskasvuun. Kasvun maksimoiva imitaation määrä on mallissa aina nolla, mutta hyvinvoinnin maksimoiva imitaation määrä voi olla positiivinen. Talouskasvun ja hyvinvoinnin maksimoivalla patentoitavissa olevan innovaation ”minimikoolla” voi olla mikä tahansa teoreettista maksimia pienempi arvo. Väitöskirjani kahdessa jälkimmäisessä pääluvussa tarkastelen informaatiohyödykkeiden kaupallista piratismia mikrotaloustieteellisen mallin avulla. Informaatiohyödykkeistä laittomasti tehtyjen kopioiden tuotantokustannukset ovat pienet, ja miltei olemattomat silloin kun niitä levitetään esimerkiksi Internetissä. Koska piraattikopioilla on monta eri tuottajaa, niiden hinnan voitaisiin mikrotaloustieteen teorian perusteella olettaa laskevan melkein nollaan, ja jos näin kävisi, kaupallinen piratismi olisi mahdotonta. Mallissani selitän kaupallisen piratismin olemassaolon olettamalla, että piratismista saatavan rangaistuksen uhka riippuu siitä, kuinka monille kuluttajille piraatti tarjoaa laittomia hyödykkeitä, ja että se siksi vaikuttaa piraattikopioiden markkinoihin mainonnan kustannuksen tavoin. Kaupallisten piraattien kiinteiden kustannusten lisääminen on mallissani aina tekijänoikeuksien haltijan etujen mukaista, mutta ”mainonnan kustannuksen” lisääminen ei välttämättä ole, vaan se saattaa myös alentaa laillisten kopioiden myynnistä saatavia voittoja. Tämä tulos poikkeaa vastaavista aiemmista tuloksista sikäli, että se pätee vaikka tarkasteltuihin informaatiohyödykkeisiin ei liittyisi verkkovaikutuksia. Aiemmin ei-kaupallisen piratismin malleista on usein johdettu tulos, jonka mukaan informaatiohyödykkeen laittomat kopiot voivat kasvattaa laillisten kopioiden myynnistä saatavia voittoja jos laillisten kopioiden arvo niiden käyttäjille riippuu siitä, kuinka monet muut kuluttajat käyttävät samanlaista hyödykettä ja jos piraattikopioiden saatavuus lisää riittävästi laillisten kopioiden arvoa. Väitöskirjan viimeisessä pääluvussa yleistän mallini verkkotoimialoille, ja tutkin yleistämäni mallin avulla sitä, missä tapauksissa vastaava tulos pätee myös kaupalliseen piratismiin.

Purification and properties of an NADP+-specific isocitrate dehydrogenase from Rhizobium meliloti

An NADP+-specific isocitrate dehydrogenase has been purified and characterized from Rhizobium meliloti. The enzyme showed Mn++ or Mg++ requirement. The apparent Km values were 2.00×10-5 m and 1.51×10-5 m for dl-isocitrate and NADP+, respectively. The enzyme was inhibited by ATP, to a lesser extent by ADP and AMP. agr-Ketoglutarate also inhibited the enzyme activity. Oxalacetate and glyoxylate together inhibited the enzyme activity. The inhibition was competitive. Studies with thiol inhibitors suggested that the enzyme contained a sulfhydryl group at or near the active site. The enzyme has an approximate molecular weight of 60 000. Fluorescence studies suggested that the enzyme contained tryptophan.

Role of Temperature in the Biological Activity of a Boreal Forest

In northern latitudes, temperature is the key factor driving the temporal scales of biological activity, namely the length of the growing season and the seasonal efficiency of photosynthesis. The formation of atmospheric concentrations of biogenic volatile organic compounds (BVOCs) are linked to the intensity of biological activity. However, interdisciplinary knowledge of the role of temperature in the biological processes related to the annual cycle and photosynthesis and atmospheric chemistry is not fully understood. The aim of this study was to improve understanding of the role of temperature in these three interlinked areas: 1) onset of growing season, 2) photosynthetic efficiency and 3) BVOC air concentrations in a boreal forest. The results present a cross-section of the role of temperature on different spatial (southern northern boreal), structural (tree forest stand - forest) and temporal (day-season- year) scales. The fundamental status of the Thermal Time model in predicting the onset of spring recovery was confirmed. However, it was recommended that sequential models would be more appropriate tools when the onset of the growing season is estimated under a warmer climate. A similar type of relationship between photosynthetic efficiency and temperature history was found in both southern and northern boreal forest stands. This result draws attention to the critical question of the seasonal efficiency of coniferous species to emit organic compounds under a warmer climate. New knowledge about the temperature dependence of the concentrations of biogenic volatile organic compounds in a boreal forest stand was obtained. The seasonal progress and the inter-correlation of BVOC concentrations in ambient air indicated a link to biological activity. Temperature was found to be the main driving factor for the concentrations. However, in addition to temperature, other factors may play a significant role here, especially when the peak concentrations are studied. There is strong evidence that the spring recovery and phenological events of many plant species have already advanced in Europe. This study does not fully support this observation. In a boreal forest, changes in the annual cycle, especially the temperature requirement in winter, would have an impact on the atmospheric BVOC composition. According to this study, more joint phenological and BVOC field observations and laboratory experiments are still needed to improve these scenarios.

Redox-linked proton transfer by cytochrome c oxidase

The respiratory chain is found in the inner mitochondrial membrane of higher organisms and in the plasma membrane of many bacteria. It consists of several membrane-spanning enzymes, which conserve the energy that is liberated from the degradation of food molecules as an electrochemical proton gradient across the membrane. The proton gradient can later be utilized by the cell for different energy requiring processes, e.g. ATP production, cellular motion or active transport of ions. The difference in proton concentration between the two sides of the membrane is a result of the translocation of protons by the enzymes of the respiratory chain, from the negatively charged (N-side) to the positively charged side (P-side) of the lipid bilayer, against the proton concentration gradient. The endergonic proton transfer is driven by the flow of electrons through the enzymes of the respiratory chain, from low redox-potential electron donors to acceptors of higher potential, and ultimately to oxygen. Cytochrome c oxidase is the last enzyme in the respiratory chain and catalyzes the reduction of dioxygen to water. The redox reaction is coupled to proton transport across the membrane by a yet unresolved mechanism. Cytochrome c oxidase has two proton-conducting pathways through which protons are taken up to the interior part of the enzyme from the N-side of the membrane. The K-pathway transfers merely substrate protons, which are consumed in the process of water formation at the catalytic site. The D-pathway transfers both substrate protons and protons that are pumped to the P-side of the membrane. This thesis focuses on the role of two conserved amino acids in proton translocation by cytochrome c oxidase, glutamate 278 and tryptophan 164. Glu278 is located at the end of the D-pathway and is thought to constitute the branching point for substrate and pumped protons. In this work, it was shown that although Glu278 has an important role in the proton transfer mechanism, its presence is not an obligatory requirement. Alternative structural solutions in the area around Glu278, much like the ones present in some distantly related heme-copper oxidases, could in the absence of Glu278 support the formation of a long hydrogen-bonded water chain through which proton transfer from the D-pathway to the catalytic site is possible. The other studied amino acid, Trp164, is hydrogen bonded to the ∆-propionate of heme a3 of the catalytic site. Mutation of this amino acid showed that it may be involved in regulation of proton access to a proton acceptor, a pump site, from which the proton later is expelled to the P-side of the membrane. The ion pair that is formed by the ∆-propionate of heme a3 and arginine 473 is likely to form a gate-like structure, which regulates proton mobility to the P-side of the membrane. The same gate may also be part of an exit path through which water molecules produced at the catalytically active site are removed towards the external side of the membrane. Time-resolved optical and electrometrical experiments with the Trp164 to phenylalanine mutant revealed a so far undetected step in the proton pumping mechanism. During the A to PR transition of the catalytic cycle, a proton is transferred from Glu278 to the pump site, located somewhere in the vicinity of the ∆-propionate of heme a3. A mechanism for proton pumping by cytochrome c oxidase is proposed on the basis of the presented results and the mechanism is discussed in relation to some relevant experimental data. A common proton pumping mechanism for all members of the heme-copper oxidase family is moreover considered.

Characterization of the molecular components and function of BARE-1, Hin-Mu and Mu transposition machineries

Transposable elements, transposons, are discrete DNA segments that are able to move or copy themselves from one locus to another within or between their host genome(s) without a requirement for DNA homology. They are abundant residents in virtually all the genomes studied, for instance, the genomic portion of TEs is approximately 3% in Saccharomyces cerevisiae, 45% in humans, and apparently more than 70% in some plant genomes such as maize and barley. Transposons plays essential role in genome evolution, in lateral transfer of antibiotic resistance genes among bacteria and in life cycle of certain viruses such as HIV-1 and bacteriophage Mu. Despite the diversity of transposable elements they all use a fundamentally similar mechanism called transpositional DNA recombination (transposition) for the movement within and between the genomes of their host organisms. The DNA breakage and joining reactions that underlie their transposition are chemically similar in virtually all known transposition systems. The similarity of the reactions is also reflected in the structure and function of the catalyzing enzymes, transposases and integrases. The transposition reactions take place within the context of a transposition machinery, which can be particularly complex, as in the case of the VLP (virus like particle) machinery of retroelements, which in vivo contains RNA or cDNA and a number of element encoded structural and catalytic proteins. Yet, the minimal core machinery required for transposition comprises a multimer of transposase or integrase proteins and their binding sites at the element DNA ends only. Although the chemistry of DNA transposition is fairly well characterized, the components and function of the transposition machinery have been investigated in detail for only a small group of elements. This work focuses on the identification, characterization, and functional studies of the molecular components of the transposition machineries of BARE-1, Hin-Mu and Mu. For BARE-1 and Hin-Mu transpositional activity has not been shown previously, whereas bacteriophage Mu is a general model of transposition. For BARE-1, which is a retroelement of barley (Hordeum vulgare), the protein and DNA components of the functional VLP machinery were identified from cell extracts. In the case of Hin-Mu, which is a Mu-like prophage in Haemophilus influenzae Rd genome, the components of the core machinery (transposase and its binding sites) were characterized and their functionality was studied by using an in vitro methodology developed for Mu. The function of Mu core machinery was studied for its ability to use various DNA substrates: Hin-Mu end specific DNA substrates and Mu end specific hairpin substrates. The hairpin processing reaction by MuA was characterized in detail. New information was gained of all three machineries. The components or their activity required for functional BARE-1 VLP machinery and retrotransposon life cycle were present in vivo and VLP-like structures could be detected. The Hin-Mu core machinery components were identified and shown to be functional. The components of the Mu and Hin-Mu core machineries were partially interchangeable, reflecting both evolutionary conservation and flexibility within the core machineries. The Mu core machinery displayed surprising flexibility in substrate usage, as it was able to utilize Hin-Mu end specific DNA substrates and to process Mu end DNA hairpin substrates. This flexibility may be evolutionarily and mechanistically important.

Reactivity of µ-Peroxo-Bridged Dimeric Vanadate in Bromoperoxidation

Diglycyl triperoxodivanadate [V2O2(O2)3(Gly H)2(H2O)2], a synthetic compound with μ-peroxo-bridge derived from H2O2and vanadate, oxidized bromide to a bromination-competent intermediate in phosphate buffer and physiological pH. This is in contrast to the requirement of acid medium with H2O2as the oxidant. Addition of its solid to bromide solution instantly produced a 262-nm-absorbing compound that converted phenol red (a trap) to its 592-nm-absorbing bromo-derivative. The high bromination activity was lost on dissolving this compound in water and the solution showed the presence of peroxovanadates (mono and di) and vanadates (V1and oligomeric V10) in51V-NMR spectrum. Of these, diperoxovanadate and vanadate together supported slow bromination activity by a second set of reactions including bromide-assisted reductive formation of vanadyl. Bromination activity dependent on vanadyl was sensitive to oxidation by excess H2O2and to complexation by EDTA, whereas that of triperoxodivanadate was relatively insensitive. Vanadyl and diperoxovanadate are capable of forming a μ-peroxo-bridged complex that is essentially similar to the synthetic vanadate dimer used in the present experiments. It appears that a μ-peroxo-intermediate is the proximal oxidant of bromide in vanadium-catalyzed bromoperoxidation.

Stereospecific photodimerization of coumarins in crystalline inclusion complexes. Molecular and crystal structure of 1:2 complex of (S,S)-(-)-1,6-bis(o-chlorophenyl)-1,6-diphenyl-hexa-2,4-diyne-1,6-diol and coumarin

Proximity of molecules is a crucial factor in many solid- state photochemical processes.'S2 The biomolecular photodimerization reactions in the solid state depend on the relative geometry of reactant molecules in the crystal lattice with center-to-center distance of nearest neighbor double bonds of the order of ca. 4 A. This fact emanates from the incisive studies of Schmidt and Cohen.2 One of the two approaches to achieve this distance requirement is the so-called "Crystal-Engineering" of structures, which essentially involves the introduction of certain functional groups that display in-plane interstacking interactions (Cl...Cl, C-He-0, etc.) in the crystal The chloro group is by far the most successful in promoting the /3- packing m ~ d e ,th~o,u~gh recent studies have shown its limitations? Another approach involves the use of constrained media in which the reactants could hopefully be aligned.