Annu Rev Plant Biol, 2002, 53, 477 - 501 A new moss genetics: targeted mutagenesis in Physcomitrella patens; Schaefer DG; The potential of moss as a model system to study plant biology is associated with their relatively simple developmental pattern that nevertheless resembles the basic organization of the body plan of land plants, the direct access to cell-lineage analysis, their similar responses to plant growth factors and environmental stimuli as those observed in other land plants, and the dominance of the gametophyte in the life cycle that facilitates genetic approaches . Transformation studies in the moss Physcomitrella patens have revealed a totally unique feature for plants, i.e., that foreign DNA sequences integrate in the genome preferentially at targeted locations by homologous recombination, enabling for the first time in plants the application of the powerful molecular genetic approaches used routinely in bacteria, yeast, and since 1989, the mouse embryonic stem cells . This article reviews our current knowledge of Physcomitrella patens transformation and its unique suitability for functional genomic studies. Planta Med, 2002 Aug, 68(8), 714 - 8 Evaluation of salicin as an antipyretic prodrug that does not cause gastric injury; Akao T et al.; Pharmacokinetic and pharmacological studies were performed to compare the antipyretic effects of salicin (SL), saligenin (SG, an aglycone of SL) and salicylic acid (SA, an active metabolite of SL) in rats . When SL was administered orally to rats, SA appeared slowly in the plasma and levels increased gradually, in contrast to the rapid appearance observed after oral administration of sodium salicylate (SANa) or SG . Orally administered SL did not affect the rectal temperatures of afebrile rats at a dose of 5 mmol/kg; at this dose, SANa and SG lowered body temperature significantly . However, it significantly reduced yeast-induced fever, producing a normal body temperature, and completely prevented fever when administered simultaneously with yeast . SL did not induce gastric lesions even at a dose of 5 mmol/kg; conversely, SANa and SG induced severe gastric lesions in a dose-dependent manner at 1, 2.5 and 5 mmol/kg . Poor absorption of SL and rapid absorption of SA and SG were confirmed in an in vivo system, as well as in an in vitro system using everted rat jejunal sacs . Only small amounts of SA and SG were detected in the intestinal tracts of rats 1 h after oral administration, whereas more than 50 % of an SL dose was recovered as SL and SG from the intestinal tracts 1 h after treatment and 15.8 % of the dose was still present as SG 4 h after administration . When given to germ-free rats, 19.8 % of the SL dose was recovered intact, mainly from the cecum, and no SG was detected even at 4 h after treatment . These results indicate that SL is a prodrug which is gradually transported to the lower part of the intestine, hydrolyzed to SG by intestinal bacteria, and converted to SA after absorption . It thus produces an antipyretic action without causing gastric injury. Spine, 2002 Sep 1, 27(17), E396 - 8 Pseudogout attack of the lumbar facet joint: a case report; Fujishiro T et al.; STUDY DESIGN: A case of acute low back pain caused by pseudogout attack of the lumbar facet joint is reported . OBJECTIVE: To report a new cause of acute low back pain previously unreported in the literature . SUMMARY OF BACKGROUND DATA: There have been some reports of lumbar spinal stenosis caused by calcium pyrophosphate dihydrate crystal deposition . However, there are no known reports of pseudogout attack of the lumbar facet joint . METHODS: An axial magnetic resonance imaging scan demonstrated joint effusion at the level of the bilateral L4-L5 facet joint . Aspiration of the left L4-L5 facet joint yielded 1.5 mL of pus-like synovial effusion . Multiple cultures of synovial fluid tested negative for bacteria and fungi, whereas compensated polarized light microscopy revealed monoclinic or triclinic crystals with a positive birefringence . RESULTS: The symptoms of acute low back pain lasted for 3 days . Local and systemic inflammatory signs, as well as symptoms, gradually improved after joint aspiration . A follow-up evaluation 8 months after lumbar facet joint aspiration showed complete resolution of pain and no neurologic deficit . CONCLUSIONS: Pseudogout attack of the lumbar facet joint is rare, but this clinical entity should be added to the differential diagnosis of acute low back pain. Proc Natl Acad Sci U S A, 2002 Sep 17, 99(19), 12236 - 41 Epub 2002 Sep 09. Experimental control of pancreatic development and maintenance; Holland AM et al.; To investigate the role of the HOX-like homeoprotein PDX1 in the formation and maintenance of the pancreas, we have genetically engineered mice so that the only source of PDX1 is a transgene that can be controlled by the application of tetracycline or its analogue doxycycline . In these mice the coding region for the tetracycline-regulated transactivator (tTA(off)) has replaced the coding region of the endogenous Pdx1 gene to ensure correct temporal and spatial expression of the regulatable transactivator . In the absence of doxycycline, tTA(off) activates the transcription of a bicistronic transgene encoding PDX1 and an enhanced green fluorescent protein reporter, which acts as a visual marker of transgene expression in living cells . Expression of the transgene-encoded PDX1 rescues the Pdx1-null phenotype; the pancreata of these mice develop and function normally . The rescue is conditional; doxycycline-mediated repression of the transgenic Pdx1 throughout gestation recapitulates the Pdx1 null phenotype . Moreover, application of doxycycline at mid-pancreogenesis blocks further development . Adult animals of the rescue genotype that were treated with doxycycline for 3 weeks shut off Pdx1 expression, decreased insulin production, and lost the ability to maintain glucose homeostasis . These results demonstrate the feasibility of controlling the formation of an organ during embryogenesis in utero and the maintenance of the mature organ through the experimental manipulation of a key developmental regulator. Trends Biotechnol, 2002 Oct, 20(10), 433 - 7 Optimizing lipases and related enzymes for efficient application; Bornscheuer UT et al.; Although numerous reactions have been performed using lipases and related enzymes (e.g . esterases and phospholipases), it is still a challenge to identify the most suitable biocatalyst and best reaction conditions for an efficient application . Frequently used methods such as immobilization and optimization of the reaction medium cannot be transferred from one reaction system or substrate to another . However, in the past few years, rational protein design and directed evolution have emerged as efficient alternative methods to optimize biocatalytic reactions. Trends Biotechnol, 2002 Oct, 20(10), 420 - 5 Trehalose production: exploiting novel approaches; Schiraldi C et al.; Trehalose (alpha-D-glucopyranosyl alpha-D-glucopyranoside) is a unique sugar capable of protecting biomolecules against environmental stress . It is a stable, colorless, odor-free and non-reducing disaccharide, and is widespread in nature . Trehalose has a key role in the survival of some plants and insects, termed anhydrobionts, in harsh environments, even when most of their water body is removed . The properties of these types of organisms drove attention towards the study of trehalose . Since then, it proved to be an active stabilizer of enzymes, proteins, biomasses, pharmaceutical preparations and even organs for transplantation . Recently, trehalose has been accepted as a safe food ingredient by the European regulation system following approval by the US Food and Drug Administration . The wide range of applications of this sugar has increased the interest of many research groups into the development of novel and economically feasible production systems . This article provides a comprehensive review of the current achievements in the biotechnological production of trehalose. Environ Microbiol, 2002 Sep, 4(9), 517 - 24 Analysis of sMMO-containing type I methanotrophs in Lake Washington sediment; Auman AJ et al.; Methane-oxidizing bacteria (methanotrophs) containing soluble methane monooxygenase (sMMO) are of interest in natural environments due to the high co-metabolic activity of this enzyme with contaminants such as trichloroethylene . We have analysed sMMO-containing methanotrophs in sediment from a freshwater lake . Environmental clone banks for a gene encoding a diagnostic sMMO subunit (mmoX) were generated using DNA extracted from Lake Washington sediment and subjected to RFLP analysis . Representatives from the six RFLP groups were cloned and sequenced, and all were found to group with Type I Methylomonas mmoX, although a majority were divergent from known Methylomonas mmoX sequences . Direct hybridization of Lake Washington sediment DNA was carried out using a series of sMMO- and Methylomonas-specific probes to assess the significance of these sMMO-containing Methylomonas-like strains in the sediment . The total sMMO-containing population and the sMMO-containing Methylomonas-like population were estimated to be similar to previous estimates for total methanotrophs and Type I methanotrophs . These results suggest that the major methanotrophic population in Lake Washington sediment consists of sMMO-containing Methylomonas-like (Type I) methanotrophs . The whole-cell TCE degradation kinetics of such a strain, LW15, isolated from this environment, were determined and found to be similar to values reported for other sMMO-containing methanotrophs . The numerical significance of sMMO-containing Methylomonas-like methanotrophs in a mesotrophic lake environment suggests that these methanotrophs may play an important role in methanotroph-mediated transformations, including co-metabolism of halogenated solvents, in natural environments. Biochemistry, 2002 Sep 17, 41(37), 11200 - 10 The two histidine axial ligands of the primary electron donor chlorophylls (P700) in photosystem I are similarly perturbed upon P700+ formation; Breton J et al.; The extent of delocalization of the positive charge in the oxidized dimer of chlorophyll (Chl) constituting P700, the primary electron donor of photosystem I (PSI), has been investigated by analyzing the perturbation upon P700(+) formation of infrared (IR) vibrational modes of the two His axial ligands of the two P700 Chl molecules . Fourier transform IR (FTIR) difference spectra of the photooxidation of P700 in PSI core complexes isolated from Synechocystis sp . PCC 6803 isotopically labeled either globally with (15)N or more specifically with (13)C on all the His residues reveal isotopic shifts of a differential signal at 1102/1108 cm(-)(1) . This signal is assigned to a downshift upon P700(+) formation of the predominantly C(5)-Ntau imidazole stretching mode of His residue(s) . The amplitude of this signal is reduced by approximately half in FTIR spectra of Synechocystis mutants in which His PsaB 651, the axial ligand to one of the two Chl molecules in P700, is replaced by Cys, Gln, or Leu . These observations provide further evidence that the positive charge in P700(+) is essentially delocalized over the two Chl molecules, in agreement with a previous FTIR study in which the frequency of the vibrational modes of the 9-keto and 10a-ester C=O groups of the two Chl's in P700, P700(+), and (3)P700 were firmly established for the first time {Breton, J., et al . (1999) Biochemistry 38, 11585-11592} . Only limited perturbations of the amplitude and frequency of the 9-keto and 10a-ester C=O bands of the P700 Chl are elicited by the mutations . On the basis of comparable mutational studies of the primary electron donor in purple bacteria, these perturbations are attributed to small molecular rearrangements of the Chl macrocycle and substituents caused by the repositioning of the P700 dimer in the new protein cavity generated by the mutations . It is proposed that the perturbation of the FTIR spectra upon mutation of a His axial ligand of the P700 Chl recently reported in Chlamydomonas reinhardtii {Hastings, G., et al . (2001) Biochemistry 40, 12943-12949} can be explained by the same effect without the need for a new assignment of the C=O bands of P700 . The distribution of charge/spin in P700(+) and (3)P700 determined by FTIR spectroscopy is discussed in relation with the contrasting interpretations derived from recent magnetic resonance experiments. Dev Biol (Basel), 2002, 108, 129 - 36 New developments in biological standardisation: commentary on the poster session on advances in transfusion safety; Seghatchian J; The 21 presentations have been arranged into two broad inter-related groups: (i) Quality of therapeutic preparations and studies on viral/bacterial inactivation procedures . (ii) Diagnostic assays for viral safety and quality control of blood components . Furthermore, in line with the main theme of this symposium, I have attempted to provide an overview of pertinent continuing investigation of blood component safety and supply, the "twin pillars" of transfusion therapy, focusing on the three pointers (3Ps) of the quality system-patient, process and product. Uisahak, 1999, 8(2), 269 - 77 {Possibility in unification of oriental and western medicine education by combination of educational curricula}; Kim JS et al.; The authors attempted a possibility of unification in the educational curricula of both Oriental and Western medical schools for the unification of two medicines . Historically the two medicines were originated from the most primitive state like instinctive method and we can say two medicines were entirely the same . However, after abrupt and current development of science in the 19th century by discovery of microscope and bacteria as well as cells, changed medicine into recent unbelievable current medicine from old ancient style medicine like Chinese Medicine which was just the remnant old medicine . The unification of educational curricula is thought to be possible to combine each other by technical adjustment from mutual understanding and cooperation for the most high quality of peoples lives . There were good equality to partial correspondences between two educational curricula around 90% at two pre-and schools from the study to analyse . The combined medicine is thought to be more efficient to the diagnosis and treatment of patients because of the effectiveness of Oriental medicine in certain disease conditions like chronic illness by acupuncture as a alternative medicine or herbs. Eur J Immunol, 2002 Sep, 32(9), 2393 - 400 Endothelin-2 is a macrophage chemoattractant: implications for macrophage distribution in tumors; Grimshaw MJ et al.; Endothelins (ET-1, ET-2 and ET-3) are 21-amino acid vasoactive peptides that bind to G-protein-linked transmembrane receptors, ET-RA and ET-RB . As well as modulating vasoconstriction, endothelins regulate growth in several cell types and may also affect differentiation, inflammation and angiogenesis . Both macrophages and endothelins are found in areas of hypoxia in solid tumors and ET-2 expression may be modulated by hypoxia in some tumors . As the peptide structure of mature endothelins is similar to that of CXC chemokines, we asked if endothelins contribute to control of macrophage distribution in tumors . We found that ET-2 is a chemoattractant for macrophages and THP-1 monocytic cells, but not for freshly isolated monocytes . The chemotactic response to ET-2 shows a typical bell-shaped response curve . Experiments with endothelin receptor antagonists showed that migration to ET-2 is mediated via the ET-RB receptor . Moreover, monocytes do not express ET-RB . Chemotaxis towards ET-2 is via the MAPK pathway: p44 and p42 are phosphorylated when THP-1 cells are stimulated with ET-2, and the MAPKK inhibitor PD98059 stops chemotaxis . As with 'classical' chemokines, migration toET-2 is also inhibited by hypoxia and by pertussis toxin . As well as its chemotactic properties, ET-2 leads to activation of macrophages . In human breast tumors that express ET-2, endothelins and ET-RB expressing macrophages often co-localized . While shorter than 'classical' chemokines, ET-2 shares a similar peptide sequence with chemokines and may signal via a similar receptor and MAPK-mediated pathway . Furthermore, ET-2 expression by tumors may modulate the behavior of macrophages such that activated cells accumulate in areas of hypoxia. Prog Nucleic Acid Res Mol Biol, 2002, 72, 165 - 221 Jasmonates and octadecanoids: signals in plant stress responses and development; Wasternack C et al.; Plants are sessile organisms . Consequently they have to adapt constantly to fluctuations in the environment . Some of these changes involve essential factors such as nutrients, light, and water . Plants have evolved independent systems to sense nutrients such as phosphate and nitrogen . However, many of the environmental factors may reach levels which represent stress for the plant . The fluctuations can range between moderate and unfavorable, and the factors can be of biotic or abiotic origin . Among the biotic factors influencing plant life are pathogens and herbivores . In case of bacteria and fungi, symbiotic interactions such as nitrogen-fixating nodules and mycorrhiza, respectively, may be established . In case of insects, a tritrophic interaction of herbivores, carnivores, and plants may occur mutualistically or parasitically . Among the numerous abiotic factors are low temperature, frost, heat, high light conditions, ultraviolet light, darkness, oxidation stress, hypoxia, wind, touch, nutrient imbalance, salt stress, osmotic adjustment, water deficit, and desiccation . In the last decade jasmonates were recognized as being signals in plant responses to most of these biotic and abiotic factors . Signaling via jasmonates was found to occur intracellularly, intercellularly, and systemically as well as interorganismically . Jasmonates are a group of ubiquitously occurring plant growth regulators originally found as the major constituents in the etheric oil of jasmine, and were first suggested to play a role in senescence due to a strong senescence-promoting effect . Subsequently, numerous developmental processes were described in which jasmonates exhibited hormone-like properties . Recent knowledge is reviewed here on jasmonates and their precursors, the octadecanoids . After discussing occurrence and biosynthesis, emphasis is placed upon the signal transduction pathways in plant stress responses in which jasmonates act as a signal . Finally, examples are described on the role of jasmonates in developmental processes. Life Sci Space Res, 1971, 9, 75 - 80 Theoretical and experimental decisions in the creation of an artificial ecosystem for human life support in space; Kirensky LV et al.; All of man's former space flights were not real ventures into space in the biological sense, as his life was supported with unregenerated earth supplies . The coming stage of space exploration requires man's long existence in the cosmos and on the other planets . This stage of man's activity outside the earth become possible only by creating small man-made ecosystems, permitting the support of his metabolism by the recycling of substances of the terrestrial biosphere . Creation of such systems is a new scientific and technical task . Man-made ecosystems are a new product of man's activity, which have no complete analogy, either in nature, or in technology . Stochastic mechanisms, which stabilize biogeocenosis, cannot be effective in small ecosystems . A technique of parametric control over biosynthesis made it possible to calculate, and put to practice, an ecosystem for man with a cyclic regeneration of the atmosphere, water and, partially, food . The specific bio-technological properties of small man-made ecosystems are being analysed . The possibility of their application for man's excursions into space and for the settlement of other planets is being considered. Life Sci Space Res, 1971, 9, 169 - 72 The influence of ultra-high vacuum on crystalline enzymes; Lysenko SV et al.; The exploration of physical conditions in space is closely related to the study of the influence of space factors on biological objects . The study of high vacuum is of particular importance because its effects are as yet imperfectly understood . Micro-organisms are suitable models for these investigations . It has already been found that in reference to high vacuum, micro-organisms may be divided into two groups: resistant and nonresistant forms . Bacterial and actinomycetal spores and conidia of Aspergillus are classified as the resistant forms . Parallel with these, rod-shaped non-sporeforming bacteria, yeasts and conidia of Penicillium, were found to be non-resistant forms . The mechanism of resistance and death of micro-organisms under high vacuum is still not clearly understood . Because of this the investigation employing such important macromolecules as crystalline cytochrome C, catalase, peroxidase and ATP have been carried out . These compounds were exposed to a high vacuum (10(-8)-10(-9) mm Hg) over a period of 72 hours . Then the weighed samples taken from treated specimens and controls were dissolved firstly in a phosphate buffer solution (pH=6.8-7.2) and were then diluted by this buffer up to a concentration of 10(-9) mg ml . The solutions obtained were investigated by the chemiluminescent method . It was found that high vacuum effects gave rise to a slight decrease (up 7%) in enzyme activity in concentrated solutions and to a significant inactivation (up to 30%) in diluted ones. Life Sci Space Res, 1971, 9, 159 - 65 Some potentialities of living organisms under simulated Martian conditions; Lozina-Lozinsky LK et al.; Temperature, humidity, pressure, composition of the atmosphere and radiation are the main factors conditioning life on the surface of Mars . When studying the Martian ecology, one must know the total effect of these factors . One may expect that, as a result of adaptation to low temperatures, there is a corresponding shift in the temperature optimum of enzymatic activity . Dryness is the main obstacle to active life . We suggest the presence of some soil moisture and water vapour . Moreover, there can be areas of permafrost . This minimum supply of water and periodic fluctuations of humidity may create conditions for the existence of drought-resistant organisms . Decreased atmospheric pressure alone does not affect micro-organisms, plants, protozoa and even insects . Ciliates reproduce in a flowing atmosphere of pure nitrogen containing 0.0002-0.0005% oxygen as an impurity . Protozoa may also develop in an atmosphere of 98-99% carbon dioxide mixed with 1% O2 . Therefore, even traces of oxygen in the Martian atmosphere would be sufficient for aerobic unicellular organisms . Cells and organisms on earth have acquired various ways of protection from uv light, and therefore may increase their resistance further by adaptation or selection . The resistance of some organisms to ionizing radiation is high enough to enable them to endure hard ionizing radiation of the sun . Experiments with unicellular {correction of unicellar} organisms show that the effect of short wave uv radiation depends on the intensity of visible light, long-wave solar uv radiation, temperatures, cell repair processes, and the state of cell components, i.e . whether the cell was frozen, dried or hydrated. Life Sci Space Res, 1968, 6, 94 - 9 On the interrelations between ultraviolet and visible light during their simultaneous action on the cell; Zaar EI; Beyond the Earth's atmosphere, ultraviolet (UV) light that is a component of the Sun's spectrum appears as one of the limiting factors to life in space and on the planets which have no ozone layer in their atmosphere . At early stages in the evolution of life, the Earth's atmosphere contained neither free oxygen nor the layers serving as a screen for short-wave UV light . Under such conditions, the UV radiation lasted for 8-10 hours a day . Similar radiation conditions may be found on Mars . Still, we do not know to what extent the photo-reactivating region of sun radiation (3000-5490 angstroms) can protect cells from the action of short UV light . Therefore, it will be of interest to study a prolonged action of UV radiation on living systems (protozoa, bacteria, fungi, animal and plant cells) in the presence of visible rays capable of reactivating UV damage . We subjected infusoria (Paramecium caudatum) to prolonged action of UV light (2537 angstroms) . The cells were irradiated daily for 8-16 hours with doses of 1.5-40 and 120 mkvt . A sharp reduction in the rate of division and the lethality to the infusoria were observed from the second to the fifth day of experimentation . In the light variant, UV radiation was performed simultaneously with the application of visible light (2800-15000 lux) . In such conditions, the cells kept their normal rate of division for 12 days . In the next 30 days without irradiation, no deviation from the normal rate was noted . The experimental results permit the conclusion that under certain conditions of interaction between visible and UV light, UV does not produce a deleterious effect on cell division. Life Sci Space Res, 1968, 6, 41 - 4 Problem of reproduction of food protein by autotrophic organisms in extraterrestrial conditions; Fofanov VI et al.; To supply a man during his prolonged stay in extraterrestrial space with dietary protein by storing it in space vehicles seems to be impractical . Protein reproduction could be realized at present only by biosynthesis, which is possible with the autotrophic organisms and is the most economic, universal and therefore quite possible for use in life-support systems . In the paper, experimental results are presented on the biological value of single-celled algae proteins . The results obtained suggested that algae biomass in human rations could not be used in quantities needed for turnover of the components of the space-cabin atmosphere . The main obstacles lie in a hard broken algae shell, a large quantity of pigment, "allergization" of persons with biomass included in their rations, lack of some aminoacids in the protein, as well as man's psychological negativism to algae as a food product . Use of single-celled algae as a food source in the future will depend on improvement of its technological and biological management . A possible use of hydrogen-reducing bacteria as a biological dietary protein product is discussed. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 1997, 29(1), 53 - 60 Comparison of Different Effects of Chloroacetates on Electron Transports in PS II and in the Reaction Center of Rb . sphaeroides 601; Yu H et al.; Chloroacetates displayed different effects on electron transports in the photosynthetic reaction center of purple bacteria (Rb.sphaeroides 601) and in the photosystem II (PS II) of higher plants . Decays of chlorophyII a fluorescence measured after actinic flashes show that chloroacetates inhibit the electron transport from Q(A)(-) to Q(B) (Q(B)(-)) and the equilibrium between Q(A)(-)Q(B) (Q(B)(-)) and Q(A)Q(B)(-) (Q(B)(2-)), acting on electron transport as well as proton transduction . The study on PSII electron transport indicates another inhibition site of chloroacetate at the oxidation side of PSII . Chloroacetates up to 500 mM have no inhibition on the DCPIPH(2) right curved arrow MV electron transport in the RS601's chromatophores . Dissociation constants of OP and trichloroacetates from RS601 reaction center were assessed to be 1.1x10(-3) M and from Dixon curve respectively . The differences on aspects of structures and functions between RS601 reaction center and PS II were discussed. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 1997, 29(1), 46 - 52 The Characteristics of DCPIPH(2) right curved arrow MV Electron Transport in Rb . sphaeroides 601; Zeng XH et al.; The absorption spectrum and fluorescence emission spectrum of RS601 were found to keep the typical characteristics of those of the purple nonsulfide bacteria Rb . sphaeroides . Under illumination, methyl viologen was reduced by RS601 chromatophores in the presence of DCPIPH(2) as the electron donor, setting up a standard noncyclic electron transport . o-phenanthroline with I(50) of 1.0 mM inhibited the DCPIPH(2) right curved arrow MV electron transport . Antimycin A did not inhibit the DCPIPH(2) right curved arrow MV electron transport and had no I(50) . The results suggested that the exact site where methyl viologen accepted electron should locate between the secondary electron acceptor, Q(B), and cyt b, but not at the Q(A) binding site as indicated before . The difference of electron transport between reduced sides of reaction centers of Rb . sphaeroides and Rs . rubrum was discussed. Rev Inst Med Trop Sao Paulo, 2002 Jul-Aug, 44(4), 209 - 12 Mycoplasma pneumoniae and Chlamydia pneumoniae in calcified nodules of aortic stenotic valves; Higuchi Mde L et al.; Aortic Valve Stenosis (AVS) has been explained as an atherosclerotic process of the valve as they often exhibit inflammatory changes with infiltration of macrophages, T lymphocytes and lipid infiltration . The present study investigated whether the bacteria Chlamydia pneumoniae (CP) and Mycoplasma pneumoniae (MP), detected previously in atherosclerotic plaques, are also present in AVS . Ten valves surgically removed from patients with AVS were analyzed by immunohistochemistry, in situ hybridization, and electron microscopy . The mean and standard deviation of the percentage areas occupied by CP antigens and MP - DNA were respectively 6.21 +/- 5.41 and 2.27 +/- 2.06 in calcified foci; 2.8 +/- 3.33 and 1.78+/- 3.63 in surrounding fibrotic areas, and 0.21 +/- 0.17 and 0.12 +/- 0.13 in less injured parts of the valve . There was higher amount of CP and MP in the calcified foci and in the surrounded fibrosis than in more preserved valvular regions . In conclusion, the fact that there were greater amounts of CP and MP in calcification foci of AVS favors the hypothesis that AS is not an inevitable degenerative process due to aging, but rather that it may be a response to the presence of these bacteria, similarly to the morphology detected in atherosclerosis damage. J Biol Chem, 2002 Nov 8, 277(45), 42852 - 8 Epub 2002 Sep 05. A coregulatory role for the TRAP-mediator complex in androgen receptor-mediated gene expression; Wang Q et al.; The human thyroid hormone receptor-associated protein (TRAP)-Mediator complex was originally identified as a large multimeric complex that copurifies with the thyroid hormone receptor (TR) from HeLa cells and markedly enhances TR-mediated transcription in vitro . More recent studies have implicated TRAP-Mediator as a coactivator for a broad range of nuclear hormone receptors as well as other classes of transcriptional activators . Here we present evidence that TRAP-Mediator plays a functional role in androgen receptor (AR)-mediated transcription . We show that several subunits of the complex ligand-dependently coimmunoprecipitate with AR from both prostate cancer LNCaP cells and from HeLa cells stably transfected with AR . The 220-kDa subunit of the complex (TRAP220) can contact the ligand-binding domain of AR in vitro, possibly implicating TRAP220 involvement in targeting AR to the holocomplex . Consistent with a TRAP-Mediator coactivator role, transient overexpression of the TRAP220, TRAP170, and TRAP100 subunits enhanced ligand-dependent transcription by AR in cultured cells . Finally, chromatin immunoprecipitation assays show that TRAP220 is recruited to the androgen-responsive prostate-specific antigen gene promoter in vivo in ligand-stimulated LNCaP cells . Collectively, these data suggest that TRAP-Mediator may play an important coregulatory role in AR-mediated gene expression. J Infect, 2002 Aug, 45(2), 99 - 106 Diarrhea, CD4 counts and enteric infections in a community-based cohort of HIV-infected adults in Uganda; Brink AK et al.; OBJECTIVES: To examine relationships between diarrhoea, CD4 cell counts and stool pathogens in a community-based cohort of HIV-infected adults in Uganda . PATIENTS AND METHODS: Stool specimens, obtained between October 1995 and December 1997, were linked to patients' symptoms and laboratory results . The relationship between CD4 counts and symptoms was tested using the Wilcoxon rank-sum test and those between organisms and diarrhoea using first a univariate Mantel-Haenszel analysis and then a logistic regression model adjusted for CD4 count and multiple organisms . RESULTS: 1,213 HIV-infected individuals (70% women, median CD4 cell count at enrollment 215 cells/microl) were followed for 1,224 person years of observation (pyo) . 484 stool samples were examined, 357 from patients with diarrhoea . The rate of diarrhoea was 661 episodes per 1,000 pyo . CD4 counts were significantly lower in individuals with diarrhoea than those without (P < 0.001, Wilcoxon rank-sum test) . Forty-nine percent of diarrhoeal stools and 39% of stools from asymptomatic patients contained enteric pathogens . The most frequent isolates were helminths (29.5% of all stools), followed by bacteria (19.2%) and then protozoa (8.9%) . Rates of isolation of diarrhoea-associated pathogens were 29% from diarrhoeal stools and 17% from asymptomatic stools (P = 0.01, chi(2) test) . The association between diarrhoea and infection with bacteria or protozoa was weak and there was no association with helminths.Cryptosporidium parvum infection alone was associated with low CD4 counts . CONCLUSIONS: Diarrhoea was common and most strongly associated with low CD4 counts . Bacteria were frequently found, even in stools from asymptomatic individuals . Over two-thirds of diarrhoeal episodes were undiagnosed, suggesting that unidentified agents or primary HIV enteropathy are important causes of diarrhoea in this population. Methods, 2002 Aug, 27(4), 324 - 32 Use of fluorescence resonance energy transfer to analyze oligomerization of G-protein-coupled receptors expressed in yeast; Overton MC et al.; Oligomerization or dimerization of G-protein-coupled receptors (GPCRs) has emerged as an important theme in signal transduction . This concept has recently gained widespread interest due to the application of direct and noninvasive biophysical techniques such as fluorescence resonance energy transfer (FRET), which have shown unequivocally that several types of GPCR can form dimers or oligomers in living cells . Current challenges are to determine which GPCRs can self-associate and/or interact with other GPCRs, to define the molecular principles that govern these specific interactions, and to establish which aspects of GPCR function require oligomerization . Although these questions ultimately must be addressed by using GPCRs expressed endogenously in their native cell types, analysis of GPCR oligomerization in heterologous expression systems will be useful to survey which GPCRs can interact, to conduct structure-function studies, and to identify peptides or small molecules that disrupt GPCR oligomerization and function . Here, we describe methods employing scanning fluorometry to detect FRET between GPCRs tagged with enhanced cyan and yellow fluorescent proteins (CFP and YFP) in living yeast cells . This approach provides a powerful means to analyze oligomerization of a variety of GPCRs that can be expressed in yeast, such as adrenergic, adenosine, C5a, muscarinic acetylcholine, vasopressin, opioid, and somatostatin receptors. Trends Biochem Sci, 2002 Sep, 27(9), 474 - 82 A universal role for MyD88 in TLR/IL-1R-mediated signaling; Janssens S et al.; The MyD88 adapter protein links members of the toll-like receptor (TLR) and interleukin-1 receptor (IL-1R) superfamily to the downstream activation of nuclear factor-kappaB and mitogen-activated protein kinases . Although originally identified as a myeloid-differentiation marker, MyD88 is now known to play an essential role in the innate immune response of insects and mammals . The generation of MyD88-deficient mice, as well as the identification of MyD88-related proteins and regulators of MyD88 signaling, has revealed new and important insights into the function of MyD88. Trends Endocrinol Metab, 2002 Oct, 13(8), 355 - 60 Aquaporins and disease: lessons from mice to humans; King LS et al.; Recent discovery of a family of water-specific membrane channel proteins, the aquaporins, has provided new insights into the molecular basis of membrane water permeability . Eleven mammalian aquaporins have been identified to date, with homolog present across the spectrum of life, including bacteria, yeast and plants . The distribution of the mammalian aquaporins predicts their participation in a range of pathophysiological events . Empirical evidence of a physiological role for aquaporins is emerging from studies in both mice and humans, and suggests that aquaporins are likely to play significant roles in human pathophysiology. Biochem J, 2002 Dec 15, 368(Pt 3), 777 - 81 The thylakoid membrane protein ALB3 associates with the cpSecY-translocase in Arabidopsis thaliana; Klostermann E et al.; The integration of light-harvesting chlorophyll proteins (LHCPs) into the thylakoid membrane requires the integral thylakoid membrane protein ALB3, a homologue of the bacterial cytoplasmic membrane protein YidC . In bacteria, YidC is associated with the SecY-translocase and facilitates the integration of Sec-dependent proteins into the plasma membrane . In addition, it is also involved in the insertion of Sec-independent proteins . In the present study we demonstrate, in Arabidopsis thaliana, that most ALB3 is a constituent of an oligomeric complex of approx . 180 kDa . In addition, we detected ALB3 in several higher-molecular-mass complexes (up to 700 kDa) . Furthermore, we show that most ALB3 co-fractionates with cpSecY during gel-filtration analysis and blue native gel electrophoresis, suggesting an association of ALB3 with the cpSecY complex . A direct interaction of ALB3 with the cpSecY complex was demonstrated by co-immunoprecipitation experiments using digitonin-solubilized thylakoid membrane proteins and anti-cpSecY or anti-ALB3 antibodies . This result was further confirmed by electron microscopic co-immunolocalization of ALB3 and cpSecY . In addition, an association of ALB3 with the cpSecY complex was demonstrated directly by cross-linking experiments using the chemical cross-linker disuccinimidyl suberate. Water Sci Technol, 2002, 46(1-2), 423 - 30 Decay rate variability of active heterotrophic biomass; Lavallee B et al.; Hypotheses of the ASM 1 model state that the specific respiration rate of heterotrophic active biomass is a constant . It thus becomes possible to assess the active heterotrophic biomass decay rate (bH) during starvation . The object of this study is to verify if the specific respiration rate of heterotrophic biomass remains constant throughout starvation, and if it is typical of global metabolic biomass activity, as proposed by the model hypotheses . The decay rate bH was evaluated using respirometric and enzymatic measurements . Measurement of bH through respirometry on various sludges varied between 0.28 and 0.76 d(-1) . According to the enzyme and respirometric measurements carried out, a large portion of this value is related to enzymatic regulation . During starvation, when initial active biomass activity is high, down regulation of the respiratory chain results in a decrease in the specific heterotrophic active biomass respiration rate, thus creating an overevaluation of the decay rate as measured by respirometry . Thus a high decay rate used in the ASM 1 model could eventually lead to a poor forecast of biomass production. Sci Prog, 2002, 85(Pt 2), 175 - 97 A new method for measuring intramolecular charge transfer; Schmuttenmaer CA; The direct measurement of intramolecular electron transfer through detection of the electromagnetic (EM) waveform that is emitted during this process is reviewed . The waveform is detected in the time-domain via free-space electro-optic sampling and then related to the dynamics of the charge transfer event . Electromagnetic generation from two systems, Betaine-30 in chloroform and DMANS in toluene, are studied to illustrate this technique . A finite-difference time-domain calculation with a time-dependent polarization is used to model the EM generation and propagation through the solution . This method is very general since the movement of charge itself generates the EM waveform, and is sensitive to charge transfer occurring on a 0.1-10 ps timescale . The potential for studying the primary steps of charge transfer in photosynthetic bacteria is also discussed. Extremophiles, 2002 Aug, 6(4), 309 - 18 Epub 2002 Mar 27. New isolates and physiological properties of the Aquificales and description of Thermocrinis albus sp . nov; Eder W et al.; The ecology of the Aquificales was studied using a combination of phylogenetic and cultivation approaches . Enrichment cultures were prepared from low-salt and marine samples of geothermally and volcanically heated environments of the United States (Yellowstone National Park), Russia (Kamchatka), Italy, Germany, Djibouti, Iceland, and Africa (Lake Tanganyika) . Isolation of single cells using the selected cell cultivation technique resulted in 15 different pure cultures . Comparisons of their 16S rRNA gene sequences showed that most of the isolates were new representatives of the major lineages of the Aquificaceae, represented by the genera Aquifex, Thermocrinis, Hydrogenobaculum, and Hydrogenobacter . Isolate HI 11/12, which was obtained from whitish streamers in the Hveragerthi area of Iceland, represents a separate branch within the Aquificaceae . The organism grew at salinities up to 0.7% NaCl and at temperatures up to 89 degrees C . Depending on the culture conditions, the organisms occurred as single motile rods, as aggregates, or as long filaments that formed whitish streamer-like cell masses . The novel isolate grew chemolithoautotrophically with hydrogen, sulfur, or thiosulfate as the electron donor under microaerophilic conditions . It represents a second species within the order Thermocrinis, which we name Thermocrinis albus HI 11/12 (DSM 14484, JCM 11386). Chin Med J (Engl), 2002 Aug, 115(8), 1205 - 8 Adenoviral mediated suicide gene transfer in the treatment of pancreatic cancer; Pan X et al.; OBJECTIVE: To determine the efficacy of adenovirus mediated suicide gene transduction combined with prodrug 5-fluorocytosine (5FC) as a therapeutic protocol for pancreatic cancer . METHODS: Cytosine Deaminase(CD) gene was cloned into pAdTrack-CMV-CD, pAdTrack-CMV-CD and pAdEasy-1 were recombined in bacteria . The newly recombined adenovirus (Ad)-CD containing green fluorescent protein (GFP) were packaged and propagated in 293 cells and purified by cesium chloride gradient centrifugation . Human pancreatic carcinoma cell line-Patu8988 was infected with this virus, then 5FC was added . XTT assay was used to estimate relative numbers of viable cells . In vivo model of pancreatic cancer was established by injecting 1.0 x 10(7) Patu8988 cells subcutaneously in Balb/c nude mice . When tumors were palpable, Ad-CD was injected into each tumor and 5FC was administered . RESULTS: Positive clones were selected using endonuclease to digest the recombinants and the concentration of viral liquids containing the CD gene was 2 x 10(11) pfu /ml . Significant cytotoxic activity as shown for 5FC in the CD gene transduced 8988 cell line, while little effect was found in the nontransduced pancreatic carcinoma cells . Antitumor effect was observed in Patu8988 xenograft nude mice with in situ CD gene transduction . CONCLUSIONS: CD gene mediated by adenovirus has high infectivity and may be useful for gene therapy in pancreatic carcinoma . These data demonstrate the use of an enzyme prodrug strategy in experimental pancreatic cancer. Hua Xi Kou Qiang Yi Xue Za Zhi, 1998 May, 16(2), 103 - 4, 113 {Comparison of demineralization of different organic acid to enamel}; Liu L et al.; The rates of demineralization of 5 organic acids (mathanoic acid, formic acid, propionic acid, Lactic acid, acetic acid, mixed acid) to the bovine enamel were tested and analysed with the self-made calcium ionselective microelectrodes(Ca(2+)-ISME) basing on a neutral carriers of ETH1001 . The results showed; 1 . The difference between the rates of demineralization of formic acid and lactic acid, formic acid and propionic acid, formic acid and acetic acid, acetic acid and mixed acid, acetic acid and lactic acid, propionic acid and mixed acid, propionic acid and lactic acid, lactic acid and mixed acid were of great significance (P < 0.01); 2 . The rates of demineralization of acetic and mixed acid decreased with time, due to saturation of the solution during demineralization; 3 . Ca(2+)-ISME was of the advantages of simplicity, rapidity, sensitivity and accuracy . The results suggest that the cariogenic potential is related to different acid products of different cariogenic bacteria, and the degree of mineral saturation within solution affects the rate of demineralization. Pathophysiol Haemost Thromb, 2002, 32 Suppl 1, 1 - 4 Disease transmission by blood products: past, present and future; Murphy WG; Transfusion of blood and blood products has been associated with transmission of infectious agents . However, it is probable that blood products are currently very safe and that pooled virus-inactivated products from remunerated donors are now safer than untreated single voluntary donor components . Although the transmission events of the past and the present are reasonably well understood, reliance on a linear approach to predict safety in the future is open to criticism . Indeed, it was not possible to predict the extent or consequences of the AIDS epidemic or of hepatitis C transmission . Moreover, although variant Creutzfeldt-Jakob disease (vCJD) may not be transmitted to any large extent by transfusion of manufactured blood products, this will be due more to good fortune than good judgement - this agent could have escaped the screening, testing and eradication methods on which current confidence in blood product safety depends . Similarly, the emergence of a highly resistant non-enveloped virus, or even of another previously unrecognised disease-causing agent, could result in new threats from transfusion of blood components and products . The ecology of blood transfusion is exquisitely sensitive to variations in starting conditions, a situation typical of a chaotic rather than a linear system . Seemingly trivial events, often apparently unrelated to blood transfusion, have had enormous consequences in this field . Whatever the events that introduced simian immunodeficiency virus to humans or scrapie to cattle, they were a long way from those involved in the manufacture of blood products . In such a setting, reliance on methods that deal effectively with known threats (such as encapsulated viruses and bacteria) without adequate investigation and management of the intrinsic sensitivity to unpredictable events, leaves open the possibility of further infections emerging in the future . It is this reality that will ultimately result in the eradication of the transfusion of donor-derived blood and blood products in the developed world . In addition, all infections with a long disease-free incubation period in the host that can be transmitted in blood will eventually be over-expressed in groups that are exposed to blood either recreationally or professionally . As in the past, this could have occurred before testing or decontamination processes have been developed for emerging pathogens . Failure to be able to rely on completely risk-free donors, in both the voluntary and non-voluntary sides of the blood industry, continues to offer the potential for the transmission of infectious diseases in the future . J Alzheimers Dis, 2001 Apr, 3(2), 181 - 190 PAMP and PARL, two novel putative metalloproteases interacting with the COOH-terminus of Presenilin-1 and -2; Pellegrini L et al.; The familial Alzheimer's disease gene products, presenilin-1 and presenilin-2 (PS1 and PS2), are involved in amyloid beta-protein precursor processing (AbetaPP), Notch receptor signaling, and programmed cell death . However, the molecular mechanisms by which presenilins regulate these processes remain unknown . Clues about the function of a protein can be obtained by seeing whether it interacts with another protein of known function . Using the yeast two-hybrid system, we identified two proteins that interact and colocalize with the presenilins . One of these newly detected presenilin-interacting proteins belongs to the FtsH family of ATP-dependent proteases, and the other one belongs to Rhomboid superfamily of membrane proteins that are highly conserved in eukaryotes, archaea and bacteria . Based on the pattern of amino acid residues conservation in the Rhomboid superfamily, we hypothesize that these proteins possess a metal-dependent enzymatic, possibly protease activity . The two putative proteases interacting with presenilins could mediate specific proteolysis of membrane proteins and contribute to the network of interactions in which presenilins are involved. Biochim Biophys Acta, 2002 Sep 13, 1577(2), 175 - 90 Elongation by RNA polymerase II: structure-function relationship; Gnatt A; RNA polymerase II is the eukaryotic enzyme that transcribes all the mRNA in the cell . Complex mechanisms of transcription and its regulation underlie basic functions including differentiation and morphogenesis . Recent evidence indicates the process of RNA chain elongation as a key step in transcription control . Elongation was therefore expected and found to be linked to human diseases . For these reasons, major efforts in determining the structures of RNA polymerases from yeast and bacteria, at rest and as active enzymes, were undertaken . These studies have revealed much information regarding the processes involved in transcription . Eukaryotic RNA polymerases and their homologous bacterial counterparts are flexible enzymes with domains that separate DNA and RNA, prevent the escape of nucleic acids during transcription, allow for extended pausing or "arrest" during elongation, allow for translocation of the DNA and more . Structural studies of RNA polymerases are described below within the context of the process of transcription elongation, its regulation and function. J Vet Pharmacol Ther, 2002 Aug, 25(4), 279 - 84 The effect of josamycine on the control of ileitis in weaned piglets under field conditions; Kyriakis SC et al.; The aim of this trial was to evaluate the effect of in-feed josamycine on the control of ileitis in weaned piglets . On a farm with a previous history of ileitis outbreaks, 288 piglets at weaning age (25 +/- 2 days old) were allocated into three experimental groups, each group comprising of four pens with 24 piglets in each pen . Group one (T1) served the trial as negative control group (unmedicated), group T2 was administered josamycine at 36 mg/kg of feed and group T3 was administered josamycine at 50 mg/kg of feed . Treatments lasted for 14 days followed by an observation period of 28 days . Administration of josamycine at both inclusion levels tested had a beneficial effect compared with the negative control group, by the reduction of prevalence of diarrhoea, the enhancement of growth performance and the reduction of prevalence of Lawsonia intracellularis in the intestine, as determined either by the PCR method or by specific histopathological examinations . The beneficial effect of josamycine was more pronounced at the inclusion level of 50 mg/kg of feed. J Vet Pharmacol Ther, 2002 Aug, 25(4), 273 - 7 Distribution of orally administered trimethoprim and sulfadiazine into noninfected subcutaneous tissue chambers in adult ponies; van Duijkeren E et al.; The distribution of trimethoprim (TMP) and sulfadiazine (SDZ) into subcutaneously implanted noninfected tissue chambers was studied in healthy adult ponies . Six ponies were given an oral TMP/SDZ paste formulation at a dose of 5 mg/kg TMP and 25 mg/kg SDZ at 12 h intervals for 2 days in order to reach steady-state concentrations . Plasma concentrations and tissue chamber fluid (TCF) concentrations of both drugs were measured at regular intervals during a period commencing 24 h after the last oral administration . The peak concentration of TMP (mean +/- SD) was 2.92 +/- 0.86 microg/mL for plasma and 1.09 +/- 0.25 microg/mL for TCF . For SDZ, the mean peak concentration was 40.20 +/- 14.74 microg/mL for plasma and 23.48 +/- 5.84 microg/mL for TCF . TMP peak concentrations in plasma were reached at 3.17 +/- 03.48 h and those in TCF at 7.33 +/- 03.72 h . SDZ peak concentrations in plasma were reached at 1.83 +/- 02.04 h and those in TCF at 8.00 +/- 03.10 h . Concentrations of TMP and SDZ in TCF remained above the generally accepted breakpoint for susceptibility (0.5/9.5 for the TMP/SDZ combination) for 12 h . Therefore, in ponies oral administration of TMP/SDZ at a dose rate of 30 mg/kg given twice daily in the form of a paste should be appropriate for effective treatment of infections caused by susceptible bacteria. Transgenic Res, 2002 Aug, 11(4), 373 - 9 Overexpression of the wheat FK506-binding protein 73 (FKBP73) and the heat-induced wheat FKBP77 in transgenic wheat reveals different functions of the two isoforms; Kurek I et al.; The FK506-binding proteins (FKBPs) belong to the peptidyl prolyl cis-trans isomerase (PPIase) family, and catalyse the rotation of the peptide bond preceding a proline . They are conserved in organisms from bacteria to man . In order to understand the function of plant FKBP isoforms, we have produced transgenic wheat plants overexpressing each of the two wheat FKBPs: wFKBP73 (which is expressed in young vegetative and reproductive tissues under normal growth conditions) and wFKBP77 (which is induced by heat stress) . Transgenic lines overexpressing wFKBP77 at 25 degrees C showed major morphological abnormalities, specifically relating to height, leaf shape, spike morphology and sterility . In these plants, the levels of hsp90 mRNA were over two fold higher than in controls, indicating a common regulatory pathway shared between wFKBP77 and Hsp90 . Transgenic lines overexpressing wFKBP73 showed normal vegetative morphology, but the grain weight and composition was altered, corresponding to changes in amylase activity during seed development. J Environ Sci (China), 2002 Jul, 14(3), 388 - 92 Photo-assisted fenton oxidation of refractory organics in UASB-pretreated leachate; Lau IW et al.; Nearly 91% of organic pollutants in Hong Kong leachate could be effectively removed by the UASB (upflow anaerobic sludge blanket) process followed by the fenton coagulation . The COD (chemical oxygen demand) of leachate was lowered from an average of 5620 mg/L to 1910 mg/L after the UASB treatment at 37 degrees C, and was further lowered to 513 mg/L after fenton coagulation . The remaining refractory residues could be further removed by photochemical oxidation with the addition of H2O2 . The BOD/COD ratio was greatly increased from 0.062 to 0.142, indicating the biodegradability of organic residues was improved . The photochemical oxidation for the fenton-coagulation supernatant was most effective at pH 3-4, with the addition of 800 mg/L of H2O2, and UV radiation time of 30 minutes . The final effluent contained only 148 mg/L of COD, 21 mg/L of BOD (biochemical oxygen demand) and 56 mg/L of TOC (total organic carbon). J Periodontol, 2002 Aug, 73(8), 886 - 91 Eikenella corrodens in subgingival plaque: relationship to age and periodontal condition; Suda R et al.; BACKGROUND: The purpose of this study was to determine the prevalence and distribution of Eikenella corrodens (E . corrodens) in subgingival plaque in different age and periodontitis groups and to examine whether its presence is related to periodontal diseases . METHODS: A total of 273 subgingival plaque samples from 213 periodontitis patients and 60 healthy subjects were assessed . Smears from each plaque sample were made and E . corrodens was detected by means of indirect immunofluorescent technique . Mean percentage of E . corrodens per total bacteria (distribution) was calculated in each sample . The prevalence (% of positive samples) and distribution of E . corrodens were statistically analyzed based on age or diagnosis by means of Fisher's exact test and analysis of variance (ANOVA) . RESULTS: Prevalence of E . corrodens decreased by age in the healthy control group; however, prevalence did not change in periodontitis groups . Distribution of E . corrodens was highest in juvenile periodontitis (JP) (2.3 +/- 1.5%) followed by post-JP (1.7 +/- 2.1%), prepubertal periodontitis (1.4 +/- 1.1%), rapidly progressive periodontitis (0.8 +/- 0.7%), adult periodontitis (0.7 +/- 0.6%), and healthy subjects (0.3 +/- 0.3%) (ANOVA, P<0.0001) . The <20-year-old age group with periodontitis showed the highest distribution of E . corrodens (2.2 +/- 1.6%) compared to the older age groups who were either healthy or had periodontitis (ANOVA, P<0.0001) . CONCLUSIONS: Since the distribution of E . corrodens is significantly higher in JP, post-JP, and PP, E . corrodens might play an important role in the occurrence or progression of periodontitis in young patients. J Periodontol, 2002 Aug, 73(8), 877 - 85 Frequency of reactivity for Porphyromonas gingivalis and Prevotella spp . in supra- and subgingival plaques, and periodontal clinical parameters according to subject age; Tanaka S et al.; BACKGROUND: The present study was conducted to assess the association between selected clinical parameters and the distribution of Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Prevotella nigrescens (Pn), and Prevotella melaninogenica (Pm) in supra- and subgingival plaque samples measured by an immunoslot blot assay (IBA) using their monoclonal antibodies . METHODS: Plaque samples from 299 patients aged 6 to 69 randomly chosen from a group of dental outpatients were examined . Plaque index, gingival index, and probing depths were evaluated according to the criteria of positive (cell number > or = 10(6)) or negative (<106) reactivity to the 4 different monoclonal antibodies . RESULTS: An increase in probing depth in subjects exhibiting either a positive or negative reaction for the 4 test bacteria was associated with increasing age . Comparing bacteria-positive subgingival plaque samples to their corresponding bacteria-negative counterparts, we found an increased plaque index in children positive for any of the 4 bacteria; in addition, that for Pg and Pi was increased in subjects 40 to 49 years old . The gingival index increased with increasing amount of Pi and Pn, but not with Pg and Pm in those 20 to 29 years of age . The frequency of Pg reactivity in subgingival plaque was markedly enhanced in subjects older than 30 to 39 years of age, and was significantly higher than that in supragingival plaque . The frequency of Pi and Pn reactivity was significantly increased in adults aged 20 to 29 and plateaued at older ages . The frequency of Pm reactivity was relatively low and independent of subject age . CONCLUSIONS: The increase in probing depth with increasing age was not affected by the occurrence of periodontopathic bacteria . The high rate of occurrence of Pg, together with Pi and Pn, in subgingival plaque of the adult age groups was suggested to be associated with the high frequency of periodontal disease in the older age groups (above 30 to 49 years of age) . The IBA appears to be useful for the efficient and rapid detection of periodontopathic bacteria. Poult Sci, 2002 Aug, 81(8), 1132 - 5 Reduction of eggshell aerobic plate counts by ultraviolet irradiation; Chavez C et al.; The effects of 254 nm ultraviolet light (UV) radiation on aerobic plate count (APC) of egg shells were investigated . In the first experiment, eggs were exposed to UV treatment (7.35 mW/cm2) for 0, 15, 30, and 60 s . Three eggs from each treatment were aseptically collected and placed into sterile plastic bags containing 50 mL of sterile phosphate-buffered solution . Serial dilutions of the phosphate-buffered solution were plated on aerobic plate count agar and incubated at 37 C for 48 h . Exposure of eggshells to 30 and 60 s UV significantly reduced aerobic plate counts compared to untreated eggs . Exposure to 60 s of UV resulted in a 2 to 3 log10 cfu/egg APC reduction and reduced counts below detectable levels . In the second experiment, UV lights were placed in a chamber equipped with a commercial-style egg conveyor . A UV treatment of 7.5 mW/cm2 and time intervals of 0, 12, 36, and 48 s were used . Three eggs were placed consecutively on the conveyor and passed through the chamber . The center egg was selected for APC evaluation . Sample size, dilution, plating, and incubation procedures were used as described for the first experiment . A significant 1 to 2 log10 reduction in colony-forming units per egg between the eggs treated 48 s to the untreated eggs was detected . The results of these studies show that UV light treatment at high intensities and low time intervals has the potential to reduce aerobic plate counts of eggshells. Comp Med, 2002 Aug, 52(4), 332 - 41 Adoptive transfer of BALb/c mouse splenocytes reduces lesion severity and induces intestinal pathophysiologic changes in the Mycobacterium avium Subspecies paratuberculosis beige/scid mouse model; Mutwiri GK et al.; Successful immune reconstitution would enhance resistance of beige/scid mice to chronic infection with Mycobacterium avium subspecies paratuberculosis, but may cause damage to intestinal tissue . Therefore, we investigated the effect of adoptive transfer of BALB/c mouse splenocytes on lesion severity and intestinal physiology in beige/scid mice infected with M . paratuberculosis . Mice were inoculated intraperitoneally (i.p.) with M . paratuberculosis, and two weeks later were inoculated i.p . with viable spleen cells from immune-competent BALB/c mice . Mice were necropsied 12 weeks after infection when engraftment of lymphocytes, clinical disease, pathologic lesions, and intestinal electrophysiologic parameters were evaluated . Lymphocytes were rare in control beige/scid mice not inoculated with spleen cells . In contrast, high numbers of CD4+, CD8+, and B220+ lymphocytes were detected in the spleen of all beige/scid mice (n = 24) inoculated with spleen cells, indicating that adoptive transfer resulted in successful engraftment of donor lymphocytes (immune reconstitution) . Immune reconstitution of M . paratuberculosis-infected beige/ scid mice significantly reduced the severity of clinical disease and pathologic lesions, and numbers of bacteria in the liver . However, intestinal electrophysiologic parameters studied in vitro indicated that intestinal tissues from reconstituted beige/scid mice had reduced short-circuit current responses (due to reduced ion secretion) following electrical, glucose, and forskolin stimulation . These abnormal responses suggested that neural or epithelial cells in the intestine were damaged . We conclude that successful immune reconstitution of beige/scid mice enhance their resistance to M . paratuberculosis infection, but may cause pathophysiologic changes associated with intestinal inflammation. Environ Mol Mutagen, 2002, 40(1), 41 - 9 Photoclastogenicity-an improved protocol, its validation, and investigation of the photogenotoxicity of DMBA; Murli H et al.; An improved protocol was developed to detect light-induced clastogenic photoproducts in Chinese hamster ovary (CHO) cells . Dishes (60 mm) containing cells and the test material or vehicle control in 3 mL of phosphate-buffered saline were exposed to light using a SUNTEST CPS solar simulation unit . Importantly, cells were exposed at about 25 cm from the light source, thereby allowing a short exposure time of 2 min . With this exposure the assay was conducted with lids removed during the UV exposure with minimal risk of contamination . After preliminary experiments an exposure of 165.6 mJ/cm(2) UVA: 17.0 mJ/cm(2) UVB was selected for treatments with the different phototoxins . Under these exposure conditions about 10-15% aberrant cells were induced in vehicle control cultures with no or minimal cytotoxicity . The well-known photoclastogens 8-methoxypsoralen (8-MOP) and chlorpromazine (CLZ) were tested . In agreement with published data, 8-MOP and CLZ were clastogenic (lowest observed effect level, LOEL, was 0.0159 microg/mL and 1.03 microg/mL, respectively) . In the absence of UV, 8-MOP was clastogenic at a much higher concentration (LOEL 251 microg/mL without UV vs . 0.0159 microg/mL with UV) while CLZ was negative up to a toxic concentration of 35 microg/mL . 7,12-Dimethylbenz{a}anthracene (DMBA), which is photomutagenic in bacteria, was clastogenic at > or =0.005 microg/mL with UV light (without S9) and at > or =2.53 microg/mL with S9 (without UV light) . These results demonstrate the utility of the protocol for the detection of photoclastogenicity and expand the characterization of DMBA's photogenotoxic activity . Arch Insect Biochem Physiol, 2002 Sep, 51(1), 46 - 54 Eicosanoids mediate Manduca sexta cellular response to the fungal pathogen Beauveria bassiana: a role for the lipoxygenase pathway; Lord JC et al.; Many studies have documented the involvement of eicosanoids in insect cellular immune responses to bacteria . The use of the fungal pathogen Beauveria bassiana as a nodulation elicitor, with inhibition of phospholipase A(2) by dexamethasone, extends the principle to fungi . This study also provides the first evidence of involvement of the lipoxygenase (LOX) pathway rather than the cyclooxygenase (COX) pathway in synthesis of the nodulation mediating eicosanoid(s) . The LOX product, 5(S)-hydroperoxyeicosa-6E,8Z,11Z,14Z-tetraenoic acid (5-HPETE), substantially reversed nodulation inhibition caused by dexamethasone and the LOX inhibitors, caffeic acid and esculetin . The COX product, prostaglandin H(2) (PGH(2)), did not reverse the nodulation inhibition by dexamethasone or the COX inhibitor, ibuprofen . None of the inhibitors tested had a significant effect on the phagocytosis of B . bassiana blastospores in vitro . Hemocyte phenoloxidase activity was reduced by dexamethasone, esculetin, and the COX inhibitor, indomethacin . The rescue candidates 5-HPETE and PGH(2) did not reverse the inhibition. Arch Insect Biochem Physiol, 2002 Sep, 51(1), 13 - 26 Co-development of Encarsia formosa (Hymenoptera: Aphelinidae) and the greenhouse whitefly, Trialeurodes vaporariorum (Homoptera: Aleyrodidae): a histological examination; Blackburn MB et al.; Using histological techniques, we have simultaneously examined the co-development of the Aphelinid parasitoid Encarsia formosa and its host the greenhouse whitefly, Trialeurodes vaporariorum . Previously we have determined that regardless of the whitefly instar parasitized, parasitoid larvae would not molt to their final instar until the whitefly reaches its maximum dimensions . In unparasitized T . vaporariorum, this point in development corresponds to the initiation of the adult molt . In part, this study was conducted to determine the developmental state of parasitized whiteflies at the time they achieve their maximum dimensions . It was found that parasitized final instar T . vaporariorum do, in fact, undergo a final molt and that E . formosa larvae will not molt to their final instar until this has occurred . The timing of the final whitefly molt appears unaffected by parasitization . The commonly observed melanization of parasitized whiteflies appears to be a consequence of this molt . In addition, we have discovered that the adult wasp oviposits within the ventral ganglion of the whitefly, and that major organ systems of the whitefly persist very late into parasitoid development . We also report the presence of possible endosymbiotic bacteria residing in the fatbody of E . formosa. Biomed Chromatogr, 2002 Aug, 16(5), 327 - 31 Capillary electrophoresis of Cr(VI) reducer Arthrobacter oxydans; Tsibakhashvili NY et al.; Rapid and effective separation of bacteria Arthrobacter oxydans was performed using capillary electrophoresis . For optimal separation of bacteria the influence of buffer concentration, pH and applied voltage were studied . It was found that the most appropriate conditions for electrophoretic mobility measurements are as follows: applied voltage 6-14 kV; buffer concentration 5-10 mM pH 6-8 . At the stationary phase of growth there are always two main heterogeneous peaks . They are connected with the morphology of bacteria as well as with cell aggregation . The heterogeneity of samples may be explained by surface modifications of bacterial cells . Microb Ecol, 2002 Oct, 44(3), 217 - 23 Epub 2002 Sep 06. Dissolved primary production and the strength of phytoplankton- bacterioplankton coupling in contrasting marine regions; Moran XA et al.; We analyzed the strength of phytoplankton-bacterioplankton coupling by comparing the rate of particulate (PPP) and dissolved primary production (DPP) with bacterial carbon demand (BCD) in four contrasting marine regions: offshore and coastal waters of the Southern Ocean, a coastal area of the NE Atlantic, and a coastal-offshore transect in the NW Mediterranean . We measured bacterial heterotrophic production (BHP) and estimated BCD from a literature model . Average phytoplanktonic percent extracellular release {PER = DPP/(DPP + PPP)} was 18-20% in the Antarctic (offshore and coastal, respectively), 16% in the NW Mediterranean, and 7% in the NE Atlantic . A significant inverse relationship was found between PER and total system productivity with pooled data . On average BHP amounted to <5% of total primary production in all regions . However, the strength of phytoplankton-bacterioplankton coupling, estimated as the potential importance of DPP in meeting BCD, differed greatly in the four regions . DPP was highly correlated to BCD in offshore Antarctic waters and was sufficient to meet BCD . In contrast, BCD exceeded DPP and bore no significant relationship in the remaining regions . The data suggest that a strong dependence of bacteria on algal extracellular production is only expected in open-ocean environments isolated from coastal inputs of DOC. Rheumatology (Oxford), 2002 Sep, 41(9), 1017 - 20 Importance of species-specific antigens in the serodiagnosis of Chlamydia trachomatis reactive arthritis; Bas S et al.; OBJECTIVES: To determine the most sensitive and specific method of anti-Chlamydia antibody measurement for the serodiagnosis of Chlamydia trachomatis reactive arthritis . METHODS: Immunoblotting, enzyme-linked immunosorbent assays using six synthetic peptides or recombinant antigens and a microimmunofluorescence test were used to determine the presence of IgG, IgM and IgA in serum samples from 17 patients with C . trachomatis reactive arthritis . Twenty patients with other inflammatory arthropathies without evidence of urogenital C . trachomatis infection were used as controls . RESULTS: The best association of sensitivity (76%) and specificity (85%) was obtained when IgG and/or IgA reactivity to two species-specific antigens was determined . These antigens were synthetic peptides, derived from species-specific epitopes in the variable domain IV of the major outer membrane protein (MOMP) (Labsystems, Finland) and recombinant polypeptide encoded by open reading frame 3 of the plasmid (pgp3) . CONCLUSIONS: IgG and/or IgA anti-MOMP-derived peptides and anti-pgp3 could be useful for the diagnosis of probable C . trachomatis reactive arthritis. Math Biosci, 2002 Sep-Oct, 179(2), 183 - 206 Plasmid-bearing, plasmid-free organisms competing for two complementary nutrients in a chemostat; Hsu SB et al.; A model of competition for two complementary nutrients between plasmid-bearing and plasmid-free organisms in a chemostat is proposed . A rigorous mathematical analysis of the global asymptotic behavior of the model is presented . The work extends the model of competition for a single-limited nutrient studied by Stephanopoulos and Lapidus {Chem . Engng . Sci . 443 (1988) 49} and Hsu, Waltman and Wolkowicz {J . Math . Biol . 32 (1994) 731}. Med Hypotheses, 2002 Aug, 59(2), 159 - 63 Impaired inactivation of digestive proteases by deconjugated bilirubin: the possible mechanism for inflammatory bowel disease; Qin XF; Inflammatory bowel disease refers to ulcerative colitis and Crohn's disease, two gut diseases with unknown causes . The dramatic increase in the last half century and the big difference in incidence for people with the same ethnic background but living in different areas strongly suggested that environmental factors played the dominant role for these diseases . The similarity in many aspects for these two diseases suggested a common causative factor . Here I suggest the impaired inactivation of digestive proteases by deconjugated bilirubin, as the result of the inhibition of bilirubin deconjugation enzyme, beta-glucuronidase, originated from the luminal bacteria and mucosa of the gut, to be a possible mechanism for both ulcerative colitis and Crohn's diseases . I also provide evidence to suggest that saccharin could be the causative or one of the most important risk factors for inflammatory bowel disease as for its inhibition on beta-glucuronidase in the intestine. Med Hypotheses, 2002 Oct, 59(4), 411 - 5 Rheumatoid arthritis is auto-immunoreaction to collagen II in cartilage happened in synovial tissue; Wang J et al.; Rheumatoid arthritis is complex and not clear on the mechanism of pathogenesis . On the basis of analysis of the symptom and pathology of rheumatoid arthritis patients, we raised a new hypothesis . The content of the hypothesis is as follows: (A) Collagen II or collagen II-Iike substance in human cartilage is the cross-autoantigen of some infecting virus or bacteria because of the structure's similarity . (B) The inflammation in synovial tissue is auto-immunoreaction to collagen II in cartilage . (C) The proliferation and attachment of synovial tissue to the surface of cartilage is due to the chemotaxis of collagen II in cartilage for the immunocytes in synovial tissue . (D) The collagenase secreted from synovial cells and immunocytes are the direct elements in the destruction of cartilage . The fallen collagen II from cartilage is one of the most important inducer on the synovial cells and immunocytes for the production of collagenase. Vet Parasitol, 2002 Sep 10, 108(2), 153 - 61 Molecular phylogenetic analysis of Onchocerca lupi and its Wolbachia endosymbiont; Egyed Z et al.; The morphology of Onchocerca lupi, responsible for canine ocular onchocercosis, is unique within the genus . Earlier analyses of the 5S ribosomal RNA gene spacer region sequence of the parasite and the 16S ribosomal RNA gene sequence of its Wolbachia endosymbiotic bacteria (Rickettsiales) supported the morphological and biological arguments that O . lupi is a distinct species . However, the exact phylogenetic position of O . lupi and its endosymbiont could not be unambiguously determined . Herein we report analyses based on the mitochondrial cytochrome oxidase I (COI) gene of the filarial species and the Wolbachia surface protein (wsp) and the bacterial cell-cycle ftsZ genes of their wolbachiae . Our results indicate that O . lupi separated from other Onchocerca spp . early in evolution . This is in line with the previous morphological analysis demonstrating that O . lupi is an atypical Onchocerca species showing both primitive and evolved characters . The phylogenetic trees generated for the COI sequences of filariae and the wsp and ftsZ sequences of their wolbachiae were congruent with each other, which supports the hypothesis that nematodes and their Wolbachia endobacteria share a long co-evolutionary history. Biochem Biophys Res Commun, 2002 Sep 6, 296(5), 1366 - 71 Acidophilic character of yeast PID261/BUD32, a putative ancestor of eukaryotic protein kinases; Facchin S et al.; Yeast piD261/Bud32 and its homologues are present in eukaryotes and in archaea but not in bacteria and are believed to make up a primordial branch of the eukaryotic protein kinase superfamily . Here, we show that, at variance with the majority of Ser/Thr protein kinases which recognize phosphoacceptor sites specified by basic and/or proline residues, piD261 phosphorylates in vitro a number of acidic proteins and peptides, and it recognizes seryl residues specified by carboxylic side chains . These data suggest that recognition of acidic sites might have been a primordial trait of protein kinases, which was modified during evolution to cope with the increasing complexity of protein phosphorylation in eukaryotes. Mol Ecol, 2002 Sep, 11(9), 1815 - 29 The incidence and diversity of Wolbachia in gallwasps (Hymenoptera; Cynipidae) on oak; Rokas A et al.; Wolbachia bacteria infect approximately 20% of all insect species, and cause a range of alterations to host reproduction, including imposition of thelytoky . The incidence and phenotypic impact of Wolbachia remains to be established in many insect taxa, and considerable research effort is currently focused on its association with particular reproductive modes and the relative importance of the various pathways via which infection occurs . Gallwasps represent an attractive system for addressing these issues for two reasons . First, they show a diversity of reproductive modes (including arrhenotoky, thelytoky and cyclical parthenogenesis) in which the impact of Wolbachia infection can be examined . Second, they occupy two intimately linked trophic niches (gall-inducers and inquilines) between which there is potential for the horizontal exchange of Wolbachia infection . In the arrhenotokous gallwasp lineages screened to date (the herb-galling 'Aylacini' and the rose-galling Diplolepidini), Wolbachia infection always induces thelytoky . The impact of Wolbachia in other arrhenotokous clades, and in the cyclically parthenogenetic clades remains unknown . Here we use polymerase chain reaction (PCR) screening and sequence data for two Wolbachia genes (wsp and ftsZ) to examine the prevalence and incidence of Wolbachia infection in 64 species (a total of 609 individuals) in two further tribes: the arrhenotokous inquilines (tribe Synergini), and the cyclically parthenogenetic oak gallwasps (tribe Cynipini) . We ask: (i) whether Wolbachia infection has any apparent impact on host reproduction in the two tribes and (ii) whether there is any correlation between Wolbachia infection and the apparent lack of an arrhenotokous generation in many oak gallwasp life cycles . We show: (i) that Wolbachia infection is rare in the Cynipini . Infected species show no deviation from cyclical parthenogenesis, and infection is no more common in species known only from a thelytokous generation; (ii) that there is a higher incidence of infection within the arrhenotokous inquilines, and generally in gallwasp tribes without cyclical parthenogensis; (iii) all Wolbachia-positive inquiline species are known to possess males, implying either that Wolbachia infection does not result in loss of sex in this tribe or, more probably, that (as for some rose gallwasps) Wolbachia infection leads to loss of sex in specific populations; and (iv) although we find some inquilines and gall inducers to be infected with Wolbachia having the same wsp sequence, these hosts are not members of the same gall communities, arguing against frequent horizontal transmission between these two trophic groups . We suggest that exchange may be mediated by the generalist parasitoids common in oak galls. J Agric Food Chem, 2002 Sep 11, 50(19), 5490 - 5 Antioxidant effects of isorhamnetin 3,7-di-O-beta-D-glucopyranoside isolated from mustard leaf (Brassica juncea) in rats with streptozotocin-induced diabetes; Yokozawa T et al.; To investigate the effects of isorhamnetin 3,7-di-O-beta-D-glucopyranoside (isorhamnetin diglucoside), a major flavonoid compound of mustard leaf, on oxidative stress due to diabetes mellitus, in vivo and in vitro studies were carried out . Oral administration of isorhamnetin diglucoside (10 or 20 mg/kg of body weight/day for 10 days) to rats with streptozotocin-induced diabetes significantly reduced serum levels of glucose and 5-(hydroxymethyl)furfural (5-HMF), which is glycosylated with hemoglobin and is an indicator of oxidative stress . After intraperitoneal administration, isorhamnetin diglucoside did not show these activities . In addition, after oral administration, the thiobarbituric acid-reactive substance levels of serum, and liver and kidney mitochondria declined significantly compared with the control group in a dose-dependent manner, whereas after intraperitoneal administration these levels fell only slightly . On the basis of the oral and intraperitoneal results, it was hypothesized that isorhamnetin diglucoside was converted to its metabolite in vivo, and its conversion to its aglycone, isorhamnetin, by beta-glucosidase was confirmed; isorhamnetin acted as an antioxidant . Moreover, it was observed that isorhamnetin diglucoside had no effect on the 1,1-diphenyl-2-picrylhydrazyl radical, whereas isorhamnetin showed a potent antioxidant effect in vitro . In addition, intraperitoneal administration of isorhamnetin reduced serum glucose and 5-HMF levels . Furthermore, lipid peroxidation in blood, liver, and kidney associated with diabetes mellitus declined after the administration of isorhamnetin . These results suggest that isorhamnetin diglucoside is metabolized in vivo by intestinal bacteria to isorhamnetin and that isorhamnetin plays an important role as an antioxidant. Clin Diagn Lab Immunol, 2002 Sep, 9(5), 1132 - 6 Determination of the nucleotide sequences of heat shock operon groESL and the citrate synthase gene (gltA) of Anaplasma (Ehrlichia) platys for phylogenetic and diagnostic studies; Inokuma H et al.; The 1,670-bp nucleotide sequence of the heat shock operon groESL and the 1,236-bp sequence of the citrate synthase gene (gltA) of Anaplasma (Ehrlichia) platys were determined . The topology of the groEL- and gltA-based phylogenetic tree was similar to that derived from 16S rRNA gene analyses with distances . Both groESL- and gltA-based PCRs specific to A . platys were also developed based upon the alignment data. Arch Environ Contam Toxicol, 2002 Oct, 43(3), 270 - 6 Nitrates/nitrites alter human lymphocyte proliferation and cytokine production; Ustyugova IV et al.; Nitrate from drinking water is converted in the body to nitrite by bacteria in the gut . This project examined effects of nitrate/nitrite on immune functions, i.e., human lymphocyte proliferation and cytokine production . Nitrate had no effect on lymphocyte growth, but nitrite decreased proliferation . Neither inhibited fibroblast growth . In 1/3 to 2/3 of the subjects tested, sodium nitrate or nitrite decreased production of Th1 cytokines (interleukin-2, interferon-gamma, and tumor necrosis factor-beta) . Nitrate and nitrite either increased or had no effect on the production of the Th2 cytokine interleukin-10 . A Th1 immune response is associated with resistance to a variety of infectious diseases; a Th2 response is associated with disease susceptibility . Because nitrate/nitrite shifted the balance from a Th1 to a Th2 response in some individuals, exposure to these compounds may decrease these persons' responsiveness to infectious diseases . The levels of nitrate used in this study are relevant to human health because they are present in the liquid portion (nonbreastfed) of some 2-month-old infants' diets in rural Romania. Nucleic Acids Res . 2002 Sep 1;30(17):e90. Cre recombinase-mediated inversion using lox66 and lox71: method to introduce conditional point mutations into the CREB-binding protein; Zhang Z et al.; CREB-binding protein (CBP) is a multifunctional cofactor implicated in many intracellular signal transduction pathways . We aimed to investigate the involvement of CBP in the cAMP response element-binding protein (CREB)-mediated pathway . The point mutation Tyr658Ala in the CREB-binding domain (CBD) was shown to abolish the binding activity of CBP to phospho-CREB, the activated form of CREB . By using a mutant Cre/loxP recombination system, this point mutation was aimed to be generated in the mouse genome in a tissue- and time-specific manner . A targeting construct in which CBD exon 5 and inverted exon 5* containing the point mutation flanked by two mutant loxP sites (lox66 and lox71) oriented in a head-to-head position was generated . When Cre recombinase is present, the DNA flanked by the two mutant loxP sites is inverted, forming one loxP and one double mutated loxP site . As the double mutated loxP site shows low affinity for Cre recombinase, the favorable reaction leads to a product where the mutated exon 5* is placed into the position to be correctly transcribed and spliced . Inversion was observed to be complete in both bacteria and mouse embryonic stem cells . Our results indicate that this Cre- mediated inversion method is a valuable tool to introduce point mutations in the mouse genome in a regulatable manner. J Clin Microbiol, 2002 Sep, 40(9), 3463 - 9 Molecular typing of papillomatous digital dermatitis-associated Treponema isolates based on analysis of 16S-23S ribosomal DNA intergenic spacer regions; Stamm LV et al.; Papillomatous digital dermatitis (PDD), an emerging infectious disease of cattle, is characterized by painful, ulcerative foot lesions . The detection of high numbers of invasive spirochetes in PDD lesions suggests an important role for these organisms in the pathogenesis of PDD . PDD-associated spirochetes have phenotypic characteristics consistent with members of the genus TREPONEMA: Partial 16S ribosomal DNA (rDNA) sequence analysis of clonal isolates from California cattle showed that they comprise three phylotypes which cluster closely with human-associated Treponema spp . of the oral cavity (T . denticola and T . medium/T . vincentii) or genital area (T . phagedenis) . The goal of our study was to apply 16S-23S rDNA intergenic spacer region (ISR) sequence analysis to the molecular typing of U.S . PDD-associated Treponema isolates . This methodology has potentially greater discriminatory power for differentiation of closely related bacteria than 16S rDNA analysis . We PCR amplified, cloned, and sequenced the ISRs from six California PDD-associated Treponema isolates and, for comparative purposes, one strain each of T . denticola, T . medium, T . vincentii, and T . phagedenis . Two ISRs that varied in length and composition were present in all the PDD-associated Treponema isolates and in T . denticola, T . medium, and T . phagedenis . ISR1 contained a tRNA(Ala) gene, while ISR2 contained a tRNA(Ile) gene . Only a single ISR (ISR1) was identified in T . vincentii . Comparative analyses of the ISR1 and ISR2 sequences indicated that the California PDD-associated Treponema isolates comprised three phylotypes, in agreement with the results of 16S rDNA analysis . PCR amplification of the 16S-tRNA(Ile) region of ISR2 permitted rapid phylotyping of California and Iowa PDD-associated Treponema isolates based on product length polymorphisms. J Clin Microbiol, 2002 Sep, 40(9), 3364 - 73 Rapid-cycle PCR and fluorimetry for detection of mycobacteria; Lachnik J et al.; In this study we used LightCycler PCR amplification and product detection by fluorescence resonance energy transfer probes to identify mycobacteria and differentiate between Mycobacterium tuberculosis complex, Mycobacterium avium, and other nontuberculous mycobacteria . Targeting the 16S rRNA gene, three different probes specific for mycobacteria, M . tuberculosis complex, and M . avium were constructed . As few as five genome copies of target nucleic acid were detected by the probes, illustrating the high sensitivity of the system . All 33 mycobacterial species tested but none of the closely related actinomycetes and other bacteria produced a specific fluorescence signal . A specificity of 100% was also demonstrated for the M . tuberculosis complex-specific probe and the M . avium-specific probe . Within 45 min, the LightCycler method correctly detected mycobacteria and specifically identified M . tuberculosis complex and M . avium without any post-PCR sample manipulation . In view of future clinical studies, we also constructed and tested an internal control which could be used to assure successful amplification and detection of mycobacteria . Monitoring of PCR inhibition will be essential for evaluation of this system for direct detection of mycobacteria in clinical specimens . Finally, we tested our system on sputum seeded with mycobacteria and were able to detect as few as 10 organisms . At present, this system is the fastest available method for identification and differentiation of mycobacteria from culture-positive specimens and offers an excellent alternative to previously established nucleic acid amplification-based techniques for the diagnostic mycobacterial laboratory. J Clin Microbiol, 2002 Sep, 40(9), 3281 - 5 Mycobacterium microti infection (vole tuberculosis) in wild rodent populations; Cavanagh R et al.; Mycobacterium microti (vole tuberculosis) infections in small wild mammals were first described more than 60 years ago in several populations in Great Britain . Few studies of vole tuberculosis have been undertaken since then, and little is known about the relationship between M . microti isolates originating from different populations or at different times or of the prevalence of this infection in wild rodent populations, despite human cases of M . microti infections being increasingly reported . In this study, field voles (Microtus agrestis), bank voles (Clethrionomys glareolus), and wood mice (Apodemus sylvaticus) were found to be infected, with up to 8% having external tuberculous signs, in wild populations in Northumberland and Cheshire, England . Spoligotyping applied directly to the clinical material simultaneously detected and typed M . microti bacteria in skin lesions, lymph glands, and internal abcesses . IS6110 restriction fragment length polymorphism typing of cultured bacteria was used to compare these isolates with previously isolated strains from both animals and humans . This demonstrated that although the current rodent isolates were distinct from those isolated from voles in the 1930s in Great Britain, they had a high degree of similarity to these strains and were distinct from the M . microti isolates from humans, a pig, and a ferret from The Netherlands . Thus, M . microti infection seems to be widespread in wild rodent populations, but more studies are needed to understand how M . microti might be transmitted from animals to humans and to determine better the zoonotic risk posed. Biophys J, 2002 Sep, 83(3), 1567 - 77 Structural change of site-directed mutants of PYP: new dynamics during pR state; Takeshita K et al.; The energetics, protein dynamics, and diffusion coefficients of three mutants of photoactive yellow protein, R52Q, P68A, and W119G, were studied by the transient grating and pulsed laser-induced photoacoustic method . We observed a new dynamics with a lifetime of approximately 1 micro s in the transient grating signal, which is silent by the light absorption technique . This fact indicates that, after the structure change around the chromophore is completed (pR(1)), the protein part located far from the chromophore is still moving to finally create another pR (pR(2)) species, which can transform to the next intermediate, pB . Although the kinetics of pR(2)-->pB-->pG are very different depending on the mutants, the enthalpies of the first long-lived (in micro seconds, 100-micro s range) intermediate species (pR(2)) are similar and very high for all mutants . The diffusion coefficients of the parent (pG) and pB species of the mutants are also similar to that of the wild-type photoactive yellow protein . From the temperature dependence of the volume change, the difference in the thermal expansion coefficients taken as indicator of the flexibility of the structure between pG and pR(2) is measured . They are also similar to that of the wild-type photoactive yellow protein . These results suggest that the protein structures of pR(2) and pB in these mutants are globally different from that of pG, and this structural change is not altered so much by the single amino acid residue mutation . This is consistent with the partially unfolded nature of these intermediate species . On the other hand, the volume changes during pR(1)-->pR(2) are sensitive to the mutations, which may suggest that the volume change reflects a rather local character of the structure, such as the chromophore-protein interaction. Anal Biochem, 2002 Aug 15, 307(2), 191 - 201 Restriction landmark genomic scanning for DNA methylation in cancer: past, present, and future applications; Rush LJ et al.; The field of molecular biology was revolutionized by the advent of gel electrophoresis . Restriction landmark genomic scanning (RLGS) is a type of two-dimensional electrophoresis employed in the genome-wide assessment of genomic alterations . RLGS has been used to study genetic and epigenetic changes in normal tissues, primary tumors, cancer cell lines, and various organisms such as mice, rats, hamsters, bacteria, and plants . An RLGS profile displays over 2000 radiolabeled restriction landmark sites in a single assay . When conducted with methylation-sensitive restriction enzymes whose sites are preferentially located in CpG island regulatory regions, RLGS becomes a very versatile tool for the investigation of both normal and aberrant methylation patterns . Early studies performed on tumor DNA were mainly descriptive in nature, essentially a catalogue of loci that were changed to varying degrees in different tumor types . Over time, as investigators have become more proficient with RLGS and have undertaken high-throughput studies, the need for efficient cloning, imaging, and analysis systems has become paramount . Current studies focus on identifying specific genes and pathways involved in deregulated methylation in cancer . As such, RLGS analysis of tumor samples has made tremendous contributions to our understanding of the role of DNA methylation in cancer . Future directions will take advantage of the abundant genomic sequence data available to link all of the RLGS loci to genes and create biologically relevant methylation profiles of cancer . This review discusses practical considerations of using RLGS as a genome scanning tool and the past, present, and future applications in cancer biology. Water Sci Technol, 2002, 45(12), 63 - 9 Waste management systems of dairy cattle farms in Japan; Shima E et al.; Recently, the size of livestock farms in Japan has been expanding and the pollution from farm wastes has become a serious problem in rural areas . Therefore it is necessary to design treatment strategies and improve the recycling of livestock manure for sustainability of agriculture in Japan . The dairy cattle waste management systems were studied at dairy farms in Aomori prefecture and in Hokkaido, Japan . The four farms, typical for the respective regions in Japan, were investigated on the basis of the land and livestock size, housing, overall farm and waste management, type of machinery and a farm labour force . A statistical comparison was made for housing, milking and waste handling systems of dairy farms . One of the waste handling strategies was aerobic slurry treatment and land irrigation of the treated liquid fraction . Such methods began to solve some of waste management problems created since 1967 in grassland farming areas of Hokkaido . The irrigation system supplies water fertiliser and organic material to land as well as shortening the spreading times . It recycles livestock resources, increases the soil fertility and rationalizes the farm management. Water Sci Technol, 2002, 45(12), 271 - 8 Ultimate azo dye degradation in anaerobic/aerobic sequential processes; Sponza DT et al.; The treatability of Remazol Black-5 was studied in an anaerobic/aerobic sequential process . Laboratory scale UASB/CSTR reactors were used and operated at different organic loadings and HRTs to investigate the COD, color removal, and methane gas production efficiencies . The effect of both sludge age and HRT on the color and COD removal efficiencies were also investigated . The reactive dye used in this study contains the groups N=N, -SO3, S=O and is in use in some of the textile industries in Turkey . The studies were carried out in continuous mode and the effluent of the UASB reactor was used as feed in the CSTR reactor . COD removal efficiencies decreased from 56 to 27% with increases in COD loadings from 5 kg COD/m3.day and 25 kg COD/m3.day in the anaerobic UASB reactor . The color removals were 92 and 87%, respectively, for the aforementioned organic loadings . The methane percentages were found to be 50 and 76% at organic loading rates of 2.49 kg COD/m3.day and 14.8 kg COD/m3.day, respectively . 28, 42, and 90% COD removal efficiencies were obtained at sludge retention times of 1.7, 5.7 and 11 days in the aerobic CSTR reactor . Optimum sludge age was 11 days in aerobic reactor and 67 and 28% COD removal efficiencies were obtained at F/M ratios of 0.05-0.17 and 0.30-1.4 kg COD/kg MLSS.day . 90-95% color and 40-60% COD removal efficiencies were obtained depending on the applied organic loadings in the UASB reactor . The remaining COD was removed with a treatment efficiency of 85-90% in the aerobic CSTR reactor. Water Sci Technol, 2002, 45(12), 113 - 8 Anaerobic digestion of organic waste in Japan: the first demonstration plant at Kyoto City; Komatsu T et al.; Recycling of Municipal Solid Waste is vigorously promoted in Japan and the necessity of energy recovery from organic waste is increasing . An anaerobic digestion demonstration plant for organic waste in Kyoto City, Japan has been operated for about two years . Three kinds of wastes (garbage and leftovers from hotels, yard waste and used paper) mixed at various ratios are used . The plant has maintained stable operations with each mixture, generating biogas by the decomposition of VS at the rate of about 820 m3N/ton-VS. J Periodontal Res, 2002 Aug, 37(4), 268 - 72 Cytolethal distending toxin of Actinobacillus actinomycetemcomitans . Occurrence and association with periodontal disease; Tan KS et al.; The cytolethal distending toxin (Cdt) of Actinobacillus actinomycetemcomitans, a periodontal pathogen, is a newly described cytotoxin with immunosuppressive properties, capable of causing cell cycle arrest of lymphocytes . The objectives of this study were to investigate the occurrence of A . actinomycetemcomitans with the cdt genotype in the subgingival plaque of periodontitis patients and to determine the association of this bacterial genotype with periodontal disease . A total of 146 subgingival plaque samples from periodontitis patients were assayed by the PCR method using oligonucleotide primers targeting the cdt operon of A . actinomycetemcomitans . Primers targeting the leukotoxin gene A (ltxA) of A . actinomycetemcomitans was used to determine the occurrence of the bacteria in the plaque samples at baseline . At baseline, A . actinomycetemcomitans was detected in 106 out of 146 (73%) diseased sites studied . Among the 106 diseased sites found to harbor A . actinomycetemcomitans, 13 sites were positive for the bacteria with the cdt genotype (12%) . Out of the 13 positive sites, 10 sites were obtained from patients diagnosed with aggressive periodontitis (77%) . Thus, A . actinomycetemcomitans with the cdt genetic subtype has low occurrence in the subgingival plaque of periodontitis patients . However, a strong association was observed between the presence of the bacteria and aggressive forms of periodontitis . Thus, the cytotoxic and immunosuppressive properties of A . actinomycetemcomitans Cdt may function to cripple the host immunity and contribute to the pathogenesis of aggressive periodontitis. J Exp Biol, 2002 Oct, 205(Pt 19), 3055 - 66 Effects of metabolite uptake on proton-equivalent elimination by two species of deep-sea vestimentiferan tubeworm, Riftia pachyptila and Lamellibrachia cf luymesi: proton elimination is a necessary adaptation to sulfide-oxidizing chemoautotrophic symbionts; Girguis PR et al.; Intracellular symbiosis requires that the host satisfy the symbiont's metabolic requirements, including the elimination of waste products . The hydrothermal vent tubeworm Riftia pachyptila and the hydrocarbon seep worm Lamellibrachia cf luymesi are symbiotic with chemolithoautotrophic bacteria that produce sulfate and protons as end-products . In this report, we examine the relationship between symbiont metabolism and host proton equivalent elimination in R . pachyptila and L . cf luymesi, and the effects of sulfide exposure on proton-equivalent elimination by Urechis caupo, an echiuran worm that lacks intracellular symbionts (for brevity, we will hereafter refer to proton-equivalent elimination as 'proton elimination') . Proton elimination by R . pachyptila and L . cf luymesi constitutes the worms' largest mass-specific metabolite flux, and R . pachyptila proton elimination is, to our knowledge, the most rapid reported for any metazoan . Proton elimination rates by R . pachyptila and L . cf luymesi correlated primarily with the rate of sulfide oxidation . Prolonged exposure to low environmental oxygen concentrations completely inhibited the majority of proton elimination by R . pachyptila, demonstrating that proton elimination does not result primarily from anaerobic metabolism . Large and rapid increases in environmental inorganic carbon concentrations led to short-lived proton elimination by R . pachyptila, as a result of the equilibration between internal and external inorganic carbon pools . U . caupo consistently exhibited proton elimination rates 5-20 times lower than those of L . cf luymesi and R . pachyptila upon exposure to sulfide . Treatment with specific ATPase inhibitors completely inhibited a fraction of proton elimination and sulfide and inorganic carbon uptake by R . pachyptila, suggesting that proton elimination occurs in large part via K(+)/H(+)-ATPases and Na(+)/H(+)-ATPases . In the light of these results, we suggest that protons are the primary waste product of the symbioses of R . pachyptila and L . cf luymesi, and that proton elimination is driven by symbiont metabolism, and may be the largest energetic cost incurred by the worms. Appl Environ Microbiol, 2002 Sep, 68(9), 4684 - 8 Enhanced degradation of an endocrine-disrupting chemical, butyl benzyl phthalate, by Fusarium oxysporum f . sp . pisi cutinase; Kim YH et al.; Compared to yeast esterase, fungal cutinase degraded butyl benzyl phthalate (BBP) far more efficiently; i.e., almost 60% of the BBP disappeared within 7.5 h . Also, the final chemical composition significantly depended on the enzyme used . Toxicity monitoring using bioluminescent bacteria showed that butyl methyl phthalate, a major product of degradation by esterase, was an oxidative toxic hazard. Appl Environ Microbiol, 2002 Sep, 68(9), 4642 - 5 Effect of Weissella confusa strain PL9001 on the adherence and growth of Helicobacter pylori; Nam H et al.; Viable and nonviable Weissella confusa strain PL9001 inhibited the binding of Helicobacter pylori to human gastric-cell line MKN-45 cells by more than 90% . Spent culture supernatant of PL9001 rapidly decreased the viability of H . pylori, rupturing cell walls . The results suggest that PL9001 is a probiotic that can reduce the infectivity and persistence of H . pylori. Appl Environ Microbiol, 2002 Sep, 68(9), 4441 - 7 Effect of modified atmosphere composition on the metabolism of glucose by Brochothrix thermosphacta; Pin C et al.; The influence of atmosphere composition on the metabolism of Brochothrix thermosphacta was studied by analyzing the consumption of glucose and the production of ethanol, acetic and lactic acids, acetaldehyde, and diacetyl-acetoin under atmospheres containing different combinations of carbon dioxide and oxygen . When glucose was metabolized under oxygen-free atmospheres, lactic acid was one of the main end products, while under atmospheres rich in oxygen mainly acetoin-diacetyl was produced . The proportions of the total consumed glucose used for the production of acetoin (aerobic metabolism) and lactic acid (anaerobic metabolism) were used to decide whether aerobic or anaerobic metabolism predominated at a given atmosphere composition . The boundary conditions between dominantly anaerobic and aerobic metabolisms were determined by logistic regression . The metabolism of glucose by B . thermosphacta was influenced not only by the oxygen content of the atmosphere but also by the carbon dioxide content . At high CO(2) percentages, glucose metabolism remained anaerobic under greater oxygen contents. Nature, 2002 Aug 29, 418(6901), 967 - 70 Epub 2002 Aug 05. Pfiesteria shumwayae kills fish by micropredation not exotoxin secretion; Vogelbein WK et al.; Pfiesteria piscicida and P . shumwayae reportedly secrete potent exotoxins thought to cause fish lesion events, acute fish kills and human disease in mid-Atlantic USA estuaries . However, Pfiesteria toxins have never been isolated or characterized . We investigated mechanisms by which P . shumwayae kills fish using three different approaches . Here we show that larval fish bioassays conducted in tissue culture plates fitted with polycarbonate membrane inserts exhibited mortality (100%) only in treatments where fish and dinospores were in physical contact . No mortalities occurred in treatments where the membrane prevented contact between dinospores and fish . Using differential centrifugation and filtration of water from a fish-killing culture, we produced 'dinoflagellate', 'bacteria' and 'cell-free' fractions . Larval fish bioassays of these fractions resulted in mortalities (60-100% in less than 24 h) only in fractions containing live dinospores ('whole water', 'dinoflagellate'), with no mortalities in 'cell-free' or 'bacteria'-enriched fractions . Videomicrography and electron microscopy show dinospores swarming toward and attaching to skin, actively feeding, and rapidly denuding fish of epidermis . We show here that our cultures of actively fish-killing P . shumwayae do not secrete potent exotoxins; rather, fish mortality results from micropredatory feeding. Alcohol Clin Exp Res, 2002 Aug, 26(8 Suppl), 66S - 69S Estriol enhances lipopolysaccharide-induced increases in nitric oxide production by Kupffer cells via mechanisms dependent on endotoxin; Enomoto N et al.; BACKGROUND: Estriol causes sensitization of Kupffer cells to lipopolysaccharide (LPS) via mechanisms dependent on gut-derived LPS . Accordingly, this study examines the effect of estriol treatment on nitric oxide (NO) production from Kupffer cells . METHODS: Rats were given estriol (20 mg/kg body weight) intraperitoneally, and Kupffer cells were isolated 24 hr later . Some rats were treated for 4 days with 150 mg/kg/day of polymyxin B and 450 mg/kg/day of neomycin to prevent growth of intestinal bacteria, the primary source of endotoxin in the gastrointestinal tract . After addition of LPS, NO production by Kupffer cell was detected using a fluorescence indicator, DAF-2 . RESULTS: Twenty-four hours after estriol administration, LPS-induced NO production by Kupffer cells was enhanced as compared with control Kupffer cells . Sterilization of the gut with antibiotics blocked this enhancement . CONCLUSIONS: Estriol treatment in vivo enhances LPS-induced NO production in Kupffer cells. EMBO J, 2002 Sep 2, 21(17), 4420 - 8 A dynamically localized histidine kinase controls the asymmetric distribution of polar pili proteins; Viollier PH et al.; Each cell division in Caulobacter crescentus is asymmetric, yielding a swarmer cell with several polar pili and a non-piliated stalked cell . To identify factors contributing to the asymmetric biogenesis of polar pili, cytological studies of pilus assembly components were performed . We show here that the CpaC protein, which is thought to form the outer membrane pilus secretion channel, and its assembly factor, CpaE, are localized to the cell pole prior to the polymerization of the pilus filament . We demonstrate that the PleC histidine kinase, a two-component signal transduction protein shown previously to localize to the piliated cell pole before and during pilus assembly, controls the accumulation of the pilin subunit, PilA . Using an inactive form of PleC (PleCH610A) that lacks the catalytic histidine residue, we provide evidence that PleC activity is responsible for the asymmetric distribution of CpaE and itself to only one of the two cell poles . Thus, a polar signal transduction protein controls its own asymmetric location as well as that of a factor assembling a polar organelle. Aliment Pharmacol Ther, 2002 Sep, 16(9), 1591 - 602 Review article: lactose--a potential prebiotic; Szilagyi A; Lactose maldigestion, which affects a large majority of the world's population, has been mostly linked with uncomfortable symptoms . In addition, dairy consumption is variably blamed or recommended for a number of ill effects . There is, however, emerging evidence that certain lactic acid-producing bacteria, which selectively consume prebiotics, may be beneficial against some lower intestinal diseases . Lactose maldigestion and lactose should perhaps be re-evaluated as a potential provider of such a prebiotic . This historical and observational review discusses lactose and argues the opinion that it has prebiotic potential . Moreover, in maldigesters, natural ingestion or lack thereof may be relevant in the pathogenesis of diseases such as colorectal cancer and inflammatory bowel diseases. Biochem Soc Trans, 2002 Aug, 30(4), 715 - 8 Mercury transport and resistance; Brown NL et al.; Resistance to mercuric ions in bacteria is conferred by mercuric reductase, which reduces Hg(II) to Hg(0) in the cytoplasmic compartment . Specific mercuric ion transport systems exist to take up Hg(II) salts and deliver them to the active site of the reductase . This short review discusses the role of transport proteins in resistance and the mechanism of transfer of Hg(II) between the mercury-resistance proteins. Biochem Soc Trans, 2002 Aug, 30(4), 621 - 4 Controlling the reactivity of radical intermediates by coenzyme B(12)-dependent methylmalonyl-CoA mutase; Banerjee R et al.; Adenosylcobalamin or coenzyme B(12)-dependent enzymes are members of the still relatively small group of radical enzymes and catalyse 1,2-rearrangement reactions . A member of this family is methylmalonyl-CoA mutase, which catalyses the isomerization of methylmalonyl-CoA to succinyl-CoA and, unlike the others, is present in both bacteria and animals . Enzymes that catalyse some of the most chemically challenging reactions are the ones that tend to deploy radical chemistry . The use of radical intermediates in an active site lined with amino acid side chains that threaten to extinguish the reaction by presenting alternative groups for abstraction poses the conundrum of how the enzymes control their reactivity . In this review, insights into this issue that have emerged from kinetic, mutagenesis and structural studies are described for methylmalonyl-CoA mutase. Biochem Soc Trans, 2002 Aug, 30(4), 613 - 20 Biosynthesis of cobalamin (vitamin B(12)); Scott AI et al.; The biosynthesis of vitamin B(12) is summarized, emphasizing the differences observed between the aerobic and anaerobic pathways . The biosynthetic route to adenosylcobalamin from its five-carbon precursor, 5-aminolaevulinic acid, can be divided into three sections: (1) the biosynthesis of uroporphyrinogen III from 5-aminolaevulinic acid, which is common to both pathways; (2) the conversion of uroporphyrinogen III into the ring-contracted, deacylated intermediate precorrin 6 or cobalt-precorrin 6, which includes the primary differences between the two pathways; and (3) the transformation of this intermediate to form adenosylcobalamin. Biochem Soc Trans, 2002 Aug, 30(4), 579 - 84 Structure and function of glutamyl-tRNA reductase involved in 5-aminolaevulinic acid formation; Moser J et al.; In most bacteria, in archaea and in plants, the general precursor of all tetrapyrroles, 5-aminolaevulinic acid, is formed by two enzymes . The initial substrate, glutamyl-tRNA, is reduced by NADPH-dependent glutamyl-tRNA reductase to form glutamate 1-semialdehyde . The aldehyde is subsequently transaminated by glutamate-1-semialdehyde 2,1-aminomutase to yield 5-aminolaevulinic acid . The enzymic mechanism and the solved crystal structure of Methanopyrrus kandleri glutamyl-tRNA reductase are described . A pathway for metabolic channelling of the reactive aldehyde between glutamyl-tRNA reductase and the aminomutase is proposed. BioDrugs, 2002, 16(4), 241 - 59 Biological imaging for the diagnosis of inflammatory conditions; Signore A et al.; Radiopharmaceuticals used for in vivo imaging of inflammatory conditions can be conve