Mol Cell Biol, 2001 Jan, 21(1), 354 - 66 Translational and structural requirements of the early nodulin gene enod40, a short-open reading frame-containing RNA, for elicitation of a cell-specific growth response in the alfalfa root cortex; Sousa C et al.; A diversity of mRNAs containing only short open reading frames (sORF-RNAs; encoding less than 30 amino acids) have been shown to be induced in growth and differentiation processes . The early nodulin gene enod40, coding for a 0.7-kb sORF-RNA, is expressed in the nodule primordium developing in the root cortex of leguminous plants after infection by symbiotic bacteria . Ballistic microtargeting of this gene into Medicago roots induced division of cortical cells . Translation of two sORFs (I and II, 13 and 27 amino acids, respectively) present in the conserved 5' and 3' regions of enod40 was required for this biological activity . These sORFs may be translated in roots via a reinitiation mechanism . In vitro translation products starting from the ATG of sORF I were detectable by mutating enod40 to yield peptides larger than 38 amino acids . Deletion of a Medicago truncatula enod40 region between the sORFs, spanning a predicted RNA structure, did not affect their translation but resulted in significantly decreased biological activity . Our data reveal a complex regulation of enod40 action, pointing to a role of sORF-encoded peptides and structured RNA signals in developmental processes involving sORF-RNAs. J Biol Chem, 2001 Mar 16, 276(11), 8427 - 35 Epub 2000 Dec 11. The complement regulator factor H binds to the surface protein OspE of Borrelia burgdorferi; Hellwage J et al.; Spirochete bacteria of the Borrelia burgdorferi sensu lato complex cause Lyme borreliosis . The three pathogenic subspecies Borrelia garinii, Borrelia afzelii, and Borrelia burgdorferi sensu stricto differ in their disease profiles and susceptibility to complement lysis . We investigated whether complement resistance of Borreliae could be due to acquisition of the main soluble inhibitors of the alternative complement pathway, factor H and the factor H-like protein 1 . When exposed to nonimmune EDTA-plasma, the serum-resistant B . afzelii and B . burgdorferi sensu stricto strains bound factor H/factor H-like protein 1 to their surfaces . Assays with radiolabeled proteins showed that factor H bound strongly to the B . burgdorferi sensu stricto strain . To identify factor H ligands on the borrelial surface, we analyzed a panel of outer surface proteins of B . burgdorferi sensu stricto with the surface plasmon resonance technique . The outer surface lipoprotein OspE was identified as a specific ligand for factor H . Using recombinant constructs of factor H, the binding site for OspE was localized to the C-terminal short consensus repeat domains 15-20 . Specific binding of factor H to B . burgdorferi sensu stricto OspE may help the pathogen to evade complement attack and phagocytosis. J Cell Sci, 2001 Jan, 114(Pt 1), 173 - 185 A novel nucleolar G-protein conserved in eukaryotes; Park JH et al.; We describe here a novel, evolutionarily conserved set of predicted G-proteins . The founding member of this family, TbNOG1, was identified in a two-hybrid screen as a protein that interacts with NOPP44/46, a nucleolar phosphoprotein of Trypanosoma brucei . The biological relevance of the interaction was verified by co-localization and co-immunoprecipitation . TbNOG1 localized to the trypanosome nucleolus and interacted with domains of NOPP44/46 that are found in several other nucleolar proteins . Genes encoding proteins highly related to TbNOG1 are present in yeast and metazoa, and related G domains are found in bacteria . We show that NOG1 proteins in humans and Saccharomyces cerevisae are also nucleolar . The S . cerevisae NOG1 gene is essential for cell viability, and mutations in the predicted G motifs abrogate function . Together these data suggest that NOG1 may play an important role in nucleolar functions . The GTP-binding region of TbNOG1 is similar to those of Obg and DRG proteins, which, together with NOG, form a newly recognized family of G-proteins, herein named ODN . The ODN family differs significantly from other G-protein families, and shows several diagnostic sequence characteristics . All organisms appear to possess an ODN gene, pointing to the biological significance of this family of G-proteins. J Invertebr Pathol, 2000 Nov, 76(4), 249 - 56 Light and electron microscopic study of a Rickettsiella species from the cockroach Blatta orientalis; Radek R; An infection with Rickettsiella sp . was responsible for an illness causing heavy body swelling in the Oriental cockroach Blatta orientalis . Reproduction of the colony stagnated . Vacuoles with parasitic bacteria occurred mainly in the fat body, but also in nearly all other organs, such as gut epithelium, Malpighian tubules, blood cells, and ovarioles . The parasites clearly differed from the symbiotic bacteria of the genus Blattabacterium, which regularly occur in the mycetocytes of B . orientalis . The vacuoles contained four stages of Rickettsiella: (1) infectious, electron-dense, rod-like elementary bodies (mean size 300 x 145 nm); (2) an electron-dense, flat intermedium stage, called flat body (mean size 515 x 255 x 125 nm); (3) an electron-light, spherical intermedium stage, called condensing sphere (mean size 340 nm); portions of cytoplasm condensed crescent-like at the border or in the center of the cell; and (4) large, spherical, electron-light initial bodies that multiplied by binary fission (mean size 600 nm) . The initial bodies had a three-layered cell boundary, but all other stages had a five-layered cell boundary . Elementary and flat bodies contained an electron-light, oblique lamella and an oval structure with an array of ribosome-like granules, respectively . In contrast to other species of Rickettsiella, crystal formation or multiple division did not occur . The described species of Rickettsiella is different from "R . blattae," which belongs to the R . popilliae group . Instead, it shares more similarities with the R . chironomi group . To avoid confusion, it was provisionally named "R . crassificans." J Oral Sci, 2000 Sep, 42(3), 139 - 46 Topography of periodontally involved human root surfaces after different chemical treatment modalities: an in vitro scanning electron microscopic study; Okte E et al.; The significance of chemical and conservative treatments of cemental tissue proximal to periodontal pockets has been pointed out in recent years . This in vitro scanning electron microscopy (SEM) study aimed to investigate the surface effects of topical applications of 0.1% cetylpyridinium chloride (CPC) and 2% sodium lauryl sulfate (SLS) and polishing on the periodontally involved root surfaces of human teeth . Ten single-rooted teeth from 8 patients with advanced adult periodontitis were included . Following extraction, any calculus was removed with extreme care to preserve as much cementum as possible . Eighty root specimens were prepared . Fresh solutions of CPC and SLS were applied for 1, 3 and 5 minutes each to 10 segments of root cementum . A total of 20 segments formed the polished (P) and control (C) groups, respectively . The results showed that the surfaces treated with CPC or SLS differed considerably from polished and control specimens . Depending on time, the surface coating was partly or wholly removed, leaving a nodular cementum structure, uncovering a fibrillar collagen substrate and the openings of dentinal tubules . Scarce debris was present on both control and polished surfaces, whereas bacteria were observed only on the control specimens . In view of these results, further definitive in vitro and in vivo research must be done to determine the advantages of chemical treatment and its effect on periodontal regeneration. Vox Sang, 2000, 79(3), 168 - 74 An experiment with glycerol-frozen red blood cells stored at -80 degrees C for up to 37 years; Valeri CR et al.; BACKGROUND AND OBJECTIVES: Red cells frozen using 40% W/V glycerol are currently FDA approved for frozen storage at -80 degrees C for up to 10 years . MATERIALS AND METHODS: Red cells frozen with 40% W/V glycerol and stored at -80 degrees C for up to 37 years were thawed, deglycerolized, and stored at 4 degrees C for 24 h . RESULTS: Red cells frozen for up to 37 years had mean freeze-thaw-wash recovery values of 75%, less than 1% hemolysis, and normal ATP, 2,3-DPG and P50 levels, and 60% of normal RBC K(+) levels . CONCLUSIONS: Red cells frozen with 40% W/V glycerol can be stored at -80 degrees C for up to 37 years with acceptable in vitro results . Biochim Biophys Acta, 2000 Dec 15, 1529(1-3), 63 - 88 Sterol methyl transferase: enzymology and inhibition; Nes WD; Sterol C-methylations catalyzed by the (S)-adenosyl-L-methionine: Delta(24)-sterol methyl transferase (SMT) have provided the focus for study of electrophilic alkylations, a reaction type of functional importance in C-C bond formation of natural products . SMTs occur generally in nature, but do not occur in animal systems, suggesting that the difference in sterol synthetic pathways can be exploited therapeutically and in insect-plant interactions . The SMT genes from several plants and fungi have been cloned, sequenced and expressed in bacteria or yeast and bioengineered into tobacco or tomato plants . These enzymes share significant amino acid sequence similarity in the putative sterol and AdoMet binding sites . Investigations of the molecular recognition of sterol fitness and studies with stereospecifically labeled substrates as well as various sterol analogs assayed with native or mutant SMTs from fungi and plants have been carried out recently in our own and other laboratories . These analyses have led to an active-site model, referred to as the 'steric-electric plug' model, which is consistent with a non-covalent mechanism involving the intermediacy of a 24beta-methyl (or ethyl) sterol bound to the ternary complex . Despite the seeming differences between fungal and plant SMT activities the recent data indicate that a distinct SMT or family of SMTs exist in these organisms which bind and transform sterols according to a similar mechanistic plan . Vascular plants have been found to express different complements of C(1)/C(2)-activities in the form of at least three SMT isoforms . This enzyme multiplicity can be a target of regulatory control to affect phytosterol homeostasis in transgenic plants . The state of our current understanding of SMT enzymology and inhibition is presented. Biochim Biophys Acta, 2000 Dec 15, 1529(1-3), 33 - 48 Isoprenyl diphosphate synthases; Wang KC et al.; Isoprenyl diphosphate synthases catalyze consecutive condensations of isopentenyl diphosphates with allylic primer substrates to form linear backbones for all isoprenoid compounds including cholesterol . These synthases are classified according to the final chain length of their end products and the stereochemistry of the newly formed double bonds . Mutagenesis and X-ray crystallography data have uncovered the basic catalytic and chain length determination mechanisms of E-isoprenyl diphosphate synthases and shed light on their possible evolutionary course . Although much less is known about the Z-isoprenyl diphosphate synthase family, successful cloning and subsequent crystallizations in the near future will no doubt bring more insight as researchers begin to unravel the essential components and precise reaction mechanisms of this cellular machinery. Mol Biol Evol, 2000 Dec, 17(12), 1956 - 70 Evolution of two-component signal transduction; Koretke KK et al.; Two-component signal transduction (TCST) systems are the principal means for coordinating responses to environmental changes in bacteria as well as some plants, fungi, protozoa, and archaea . These systems typically consist of a receptor histidine kinase, which reacts to an extracellular signal by phosphorylating a cytoplasmic response regulator, causing a change in cellular behavior . Although several model systems, including sporulation and chemotaxis, have been extensively studied, the evolutionary relationships between specific TCST systems are not well understood, and the ancestry of the signal transduction components is unclear . Phylogenetic trees of TCST components from 14 complete and 6 partial genomes, containing 183 histidine kinases and 220 response regulators, were constructed using distance methods . The trees showed extensive congruence in the positions of 11 recognizable phylogenetic clusters . Eukaryotic sequences were found almost exclusively in one cluster, which also showed the greatest extent of domain variability in its component proteins, and archaeal sequences mainly formed species-specific clusters . Three clusters in different parts of the kinase tree contained proteins with serine-phosphorylating activity . All kinases were found to be monophyletic with respect to other members of their superfamily, such as type II topoisomerases and Hsp90 . Structural analysis further revealed significant similarity to the ATP-binding domain of eukaryotic protein kinases . TCST systems are of bacterial origin and radiated into archaea and eukaryotes by lateral gene transfer . Their components show extensive coevolution, suggesting that recombination has not been a major factor in their differentiation . Although histidine kinase activity is prevalent, serine kinases have evolved multiple times independently within this family, accompanied by a loss of the cognate response regulator(s) . The structural and functional similarity between TCST kinases and eukaryotic protein kinases raises the possibility of a distant evolutionary relationship. Mol Biol Evol, 2000 Dec, 17(12), 1807 - 15 Reconstructing genealogies of serial samples under the assumption of a molecular clock using serial-sample UPGMA; Drummond A et al.; Reconstruction of evolutionary relationships from noncontemporaneous molecular samples provides a new challenge for phylogenetic reconstruction methods . With recent biotechnological advances there has been an increase in molecular sequencing throughput, and the potential to obtain serial samples of sequences from populations, including rapidly evolving pathogens, is fast being realized . A new method called the serial-sample unweighted pair grouping method with arithmetic means (sUPGMA) is presented that reconstructs a genealogy or phylogeny of sequences sampled serially in time using a matrix of pairwise distances . The resulting tree depicts the terminal lineages of each sample ending at a different level consistent with the sample's temporal order . Since sUPGMA is a variant of UPGMA, it will perform best when sequences have evolved at a constant rate (i.e., according to a molecular clock) . On simulated data, this new method performs better than standard cluster analysis under a variety of longitudinal sampling strategies . Serial-sample UPGMA is particularly useful for analysis of longitudinal samples of viruses and bacteria, as well as ancient DNA samples, with the minimal requirement that samples of sequences be ordered in time. J Biol Chem, 2001 Mar 9, 276(10), 7437 - 41 Epub 2000 Dec 07. Direct demonstration that homotetrameric chaperone SecB undergoes a dynamic dimer-tetramer equilibrium; Topping TB et al.; We have shown here that the cytosolic bacterial chaperone SecB is a structural dimer of dimers that undergoes a dynamic equilibrium between dimer and tetramer in the native state . We demonstrated this equilibrium by mixing two tetrameric species of SecB that can be distinguished by size . We showed that the homotetrameric species exchanged dimers, because when the mixture was analyzed both by size exclusion chromatography and native polyacrylamide gel electrophoresis a third hybrid tetrameric species was detected . Furthermore, treatment of SecB with 5,5'-dithiobis-(2-nitrobenzoic acid), which modifies the sulfhydryl group on cysteines, caused irreversible dissociation to a dimer indicating that cysteine must be involved in the stabilizing interactions at the dimer interface . It is clear that the two dimer-dimer interfaces of the SecB tetramer are differentially stable . Dissociation at one interface allows for a dynamic dimer-tetramer equilibrium . Because only dimers were exchanged it is clear that the other interface between dimers is significantly more stable, otherwise oligomers should have formed with a random distribution of monomers. ASAIO J, 2000 Nov-Dec, 46(6), S2 - 5 Physiology of wound healing and surgical wound care; Phillips SJ; Wound healing is a systemic process, which occurs stepwise and involves the stages of hemostasis, inflammation, and repair . Hemostasis with fibrin formation creates a protective wound scab . The scab provides a surface beneath which cell migration and movement of the wound edges can occur . Inflammation brings nutrients to the area of the wound, removes debris and bacteria, and provides chemical stimuli for wound repair . Repair begins immediately after wounding and proceeds rapidly through the processes of epithelialization, fibroplasia, and capillary proliferation into the healing area . Different tissues have their own normal rates of growth during the process of healing . The optimal rate of healing is approached when factors advantageous to healing are present and factors having the ability to disturb or retard the healing processes are controlled or absent . These factors are discussed. Przegl Lek, 2000, 57(7-8), 424 - 6 {Tick spirochetosis--Lyme borreliosis}; Nowakowski G et al.; Lyme boreliosis is currently the most common tick-borne infection . It may cause various clinical symptoms depending on organ localization and duration of the infection . The disease may be symptomless, subclinical or with full clinical manifestation . Usually three clinical stages may be distinguished . In stage I erythema migrans and flu-like symptoms usually develop . In stage II, connected with the infection spreading with blood and lymph, beside joint pains, neuroboreliosis appears, sometimes the disease involves other organs such as heart, eyes, testicles, joints . Stage III, chronic in its character, usually develops in patients who had previously reported joint and neurological complaints . Encephalopathy and fibromyalgia accompany joint involvement . Diagnostics of Lyme borreliosis is based on clinical evaluation and laboratory test including culture of the bacteria obtained from biopsies and serological tests . There are no established standards of the treatment--some examples of the therapy are presented in the paper . The disease if not treated has a progressive course in most causes, however in some patients it can resolve spontaneously even with no treatment. J Public Health Dent, 2000 Summer, 60(3), 197 - 206; discussion 207-9 Dietary determinants of dental caries and dietary recommendations for preschool children; Tinanoff N et al.; OBJECTIVES: The purpose of this review, commissioned by the Administration for Children and Families, the Health Resources and Services Administration, the Health Care Financing Administration, and the Department of Agriculture's Food and Nutrition Service, was to update the evidence of the dietary factors that affect dental caries, and subsequently formulate dietary recommendations for preschool children based on principles of cariology . METHODS: Literature on the dental caries process, dietary factors affecting dental caries initiation and progression, and nutrition education and counseling were reviewed and synthesized . Dietary guidelines for children at various ages were then constructed based on the review . RESULTS: Dental caries in preschool children is due to a combination of factors, including colonization of teeth with cariogenic bacteria, type of foods and frequency of exposure of these foods to the cariogenic bacteria, and susceptible teeth . Caries risk is greatest if sugars are consumed at high frequency and are in a form that is retained in the mouth for long periods . Sucrose is the most cariogenic sugar because it can form glucan that enables firm bacterial adhesion to teeth and limits diffusion of acid and buffers in the plaque . There is emerging interest in the effects of tooth development and its role in the future dental caries risk of the child . CONCLUSIONS: Nutrition education and counseling for the purposes of reducing caries in children is aimed at teaching parents the importance of reducing high frequency exposures to obvious and hidden sugars . Guidelines include: avoiding frequent consumption of juice or other sugar-containing drinks in the bottle or sippy cup, discouraging the behavior of a child sleeping with a bottle, promoting noncariogenic foods for snacks, fostering eating patterns consistent with the Food Guide Pyramid, limiting cariogenic foods to mealtimes, rapidly clearing cariogenic foods from the child's oral cavity either by toothbrushing or by consumption of protective foods, and restricting sugar-containing snacks that are slowly eaten (e.g., candy, cough drops, lollipops, suckers) . Along with nutritional factors, a comprehensive approach to preventing dental caries in preschool children must include improved general dietary habits, good oral hygiene, appropriate use of fluorides, and access to preventive and restorative dental care. Diagn Microbiol Infect Dis, 2000 Nov, 38(3), 169 - 70 Feculent meningitis: polymicrobial meningitis in colorectal surgery; Garcia-Lechuz JM et al.; Polymicrobial anaerobic meningitis is a rare event secondary to a contiguous infection in the head or neck . Anaerobic meningitis due to a meningo-intestinal fistula is a rare event with only two cases reported in the literature . We describe a new case of adult polymicrobial anaerobic meningitis after colorectal surgery and radiotherapy and review the previous two cases. FEBS Lett, 2000 Dec 1, 486(1), 52 - 6 Isolation, reconstitution and functional characterisation of the Rhodobacter sphaeroides photoactive yellow protein; Haker A et al.; We report the isolation, functional reconstitution and photophysical characterisation of Rhodobacter sphaeroides photoactive yellow protein (PYP), of which the gene was recently cloned . Reconstitution of the his-tagged purified apo-protein with 4-hydroxy-cinnamic acid yields the characteristic blue absorbance at 446 nm, but surprisingly also an absorbance peak at 360 nm . This additional peak is not caused by binding of a second chromophore, as confirmed with mass spectroscopy . Moreover, reconstitution with the 'locked' analogue 7-hydroxy-coumarin-3-carboxylic acid yields only a single absorbance peak at 441 nm . The 446 nm and 360 nm species are part of a temperature- and pH-dependent equilibrium . Photoactivation of the protein leads to formation of a blue-shifted intermediate as in other PYPs, with a 100-fold increased groundstate recovery rate (k(pB-->pG)=500 s(-1)) compared to E-PYP. Int Microbiol, 2000 Mar, 3(1), 45 - 9 DNA fluorescent stain accumulates in the Golgi but not in the kinetosomes of amitochondriate protists; Dolan MF; Hindgut symbiotic trichomonads (uninucleate Caduceia versatilis, and multinucleate Stephanonympha sp . and Snyderella tabogae) from the dry-wood-eating termite Cryptotermes cavifrons (Kalotermitidae) accumulate DAPI (4,6diamidino-2-phenylindole) in the membranous sacs of the Golgi complex . This form of Golgi complex, typical of protists in the class Parabasalia, is called a parabasal body . Trichomonads contain organellar systems, mastigonts, that consist of four undulipodia (e.g . eukaryotic flagella and cilia), axostylar microtubules, a parabasal body and other structures . These cells bear from one (in the case of Caduceia) to hundreds (in the case of Snyderella) of mastigonts . These features are characteristic of their protist class (Parabasalia) . The nuclei of all three species stained with DNA-specific stains: DAPI, SYTOX, acridine orange, propidium iodide, ethidium bromide and Feulgen, at optimal concentrations, but kinetosomes failed to stain at all . The nuclei, parabasal bodies and symbiotic bacteria (but no microtubular structures) fluoresced in glutaraldehyde-fixed cells stained with 1.45 microM DAPI . Parabasal bodies of Snyderella and Caduceia treated to remove lipids with Triton X-100, or treated with 5% trichloroacetic acid, lacked DAPI-fluorescence . I conclude that DNA, present as expected in nuclei and bacterial symbionts, is absent from and not associated with calonymphid kinetosomes . The reason for DNA-RNA stain accumulation in the Golgi cistemae is not clear. Int Microbiol, 2000 Mar, 3(1), 17 - 24 Biosorption: a solution to pollution? Vieira RH, Volesky B. To solve the water pollution problem by toxic heavy metal contamination resulting from humans technological activities has for long presented a challenge . Biosorption can be a part of the solution . Some types of biosorbents such as seaweeds, molds, yeasts, bacteria or crab shells are examples of biomass tested for metal biosorption with very encouraging results . The uptake of heavy metals by biomass can in some cases reach up to 50% of the biomass dry weight . New biosorbents can be manipulated for better efficiency and multiple re-use to increase their economic attractiveness. Int Microbiol, 1999 Sep, 2(3), 155 - 60 Molecular mechanisms of malaria sporozoite motility and invasion of host cells; Sultan AA; Malaria sporozoites have the unique capacity to invade two entirely different types of target cell in the mosquito vector and the vertebrate host during the course of the parasite's life cycle . Although little is known about the specific interaction of the sporozoite with its target cells, two sporozoite proteins, circumsporozoite (CS) and thrombospondin-related adhesive protein (TRAP), have been shown to play important roles in the invasion of both cell types . CS protein is a multifunctional protein involved in sporogony, invasion of the salivary glands, the specific arrest of sporozoites in the liver sinusoid, gliding motility of the sporozoite, and hepatocyte recognition and entry . TRAP has been shown to be critical for sporozoite infection of the mosquito salivary glands and liver cells, and is essential for sporozoite gliding motility . This review will focus on the involvement of these molecules in sporozoite motility and the invasion of host cells. Int Microbiol, 1999 Sep, 2(3), 137 - 44 Molecular aspects of Bordetella pertussis pathogenesis; Locht C; The molecular mechanisms of Bordetella virulence are now well understood, and many virulence factors have been identified and characterized at the molecular level . These virulence factors can be grouped into two major categories: adhesins, such as filamentous hemagglutinin, pertactin and fimbriae, and toxins, such as pertussis toxin, adenylate cyclase, dermonecrotic toxin and tracheal cytotoxin . The production of most virulence factors is coordinately regulated by a two-component signal transduction system composed of the regulator BvgA and the sensor protein BvgS . The adhesins and toxins act in concert to establish infection . Some adhesins exert their effects synergically or are redundant functioning only in the absence of another adhesin, illustrating the importance of adhesion in infection . Most virulence factors are secreted into the culture supernatant or exposed at the surface of the bacterial cell . A notable exception is dermonecrotic toxin, which remains in the cytoplasmic compartment of bacterial cells . Most virulence factors are produced by all of the three major Bordetella species, B . pertussis, B . parapertussis and B . bronchiseptica . However, some, such as pertussis toxin and the tracheal colonization factor, are only produced by B . pertussis . Our understanding of Bordetella virulence at the molecular level has led to the development of new acellular vaccines against whooping cough, and of genetically attenuated B . pertussis strains to be used as recombinant live bacterial vaccine vectors for homologous and heterologous protection. Int Microbiol, 1998 Dec, 1(4), 319 - 26 Morphogenesis by symbiogenesis; Chapman MJ et al.; Here we review cases where initiation of morphogenesis, including the differentiation of specialized cells and tissues, has clearly evolved due to cyclical symbiont integration . For reasons of space, our examples are drawn chiefly from the plant, fungal and bacterial kingdoms . Partners live in symbioses and show unique morphological specializations that result when they directly and cyclically interact . We include here brief citations to relevant literature where plant, bacterial or fungal partners alternate independent with entirely integrated living . The independent, or at least physically unassociated stages, are correlated with the appearance of distinctive morphologies that can be traced to the simultaneous presence and strong interaction of the plant with individuals that represent different taxa. Evolution Int J Org Evolution, 2000 Oct, 54(5), 1661 - 72 Expression of cytoplasmic incompatibility in Drosophila simulans and its impact on infection frequencies and distribution of Wolbachia pipientis; James AC et al.; The aim of this study is to examine the expression of cytoplasmic incompatibility and investigate the distribution and population frequencies of Wolbachia pipientis strains in Drosophila simulans . Nucleotide sequence data from 16S rDNA and a Wolbachia surface protein coding sequence and cytoplasmic incompatibility assays identify four distinct Wolbachia strains: wHa, wRi, wMa, and wAu . The levels of cytoplasmic incompatibility between six lines carrying these strains of bacteria and three control lines without bacteria are characterized . Flies infected with wHa and wRi are bidirectionally incompatible, and males that carry either strain can only successfully produce normal numbers of offspring with females carrying the same bacterial strain . Males infected with wAu do not express incompatibility . Males infected with the wMa strain express intermediate incompatibility when mated to females with no bacteria and no incompatibility with females with any other Wolbachia strain . We conduct polymerase chain reaction/restriction fragment length polymorphism assays to distinguish the strain of Wolbachia and the mitochondrial haplotype to survey populations for each type and associations between them . Drosophila simulans is known to have three major mitochondrial haplotypes (siI, sill, and siIII) and two subtypes (siIIA and siIIB) . All infected lines of the sil haplotype carry wHa, wNo, or both; wMa and wNo are closely related and it is not clear whether they are distinct strains or variants of the same strain . Infected lines with the silIA haplotype harbor wRi and the siIIB haplotype carries wAu . The wMa infection is found in siIII haplotype lines . The phenotypic expression of cytoplasmic incompatibility and its relation to between-population differences in frequencies of Wolbachia infection are discussed. Cytokines Cell Mol Ther, 2000 Jun, 6(2), 89 - 95 Use of G-CSF for granulocyte transfusion therapy; Hubel K et al.; Patients with neutropenia, especially neutropenia following aggressive myeloablative therapy, are at high risk for developing infectious complications caused by bacteria and opportunistic fungi . Infections remain one of the leading causes of treatment failure in patients with cancer . Thus, new and innovative therapeutic strategies are needed for management of neutropenic patients with infection . Because neutrophils represent the first line of host defense, granulocyte transfusion therapy should be a logical therapeutic approach . Although such therapy has been employed sporadically for several decades, clinical benefit has been compromised by technical problems and low granulocyte yields resulting from inadequate donor stimulation . The discovery of granulocyte colony-stimulating factor (G-CSF) as a means to elevate blood neutrophil counts when administered to normal donors has rekindled interest in granulocyte transfusion therapy . Extensive experience has been gained worldwide with G-CSF in clinical practice, and adverse events have been minimal when G-CSF has been administered to patients or healthy persons in human trials . This review focuses on the use of G-CSF in granulocyte transfusion therapy, including technical considerations of granulocyte leukapheresis and storage, donor selection and stimulation, as well as treatment results and associated risks. Angiology, 2000 Oct, 51(10), 867 - 71 Abdominal aortic aneurysm infected with Helicobacter pylori--a case report; Hirose H et al.; The authors present a case of an abdominal aortic aneurysm infected with Helicobacter pylori bacteria . In their literature search, the authors found no other report of the Helicobacter pylori involved in an infected abdominal aortic aneurysm. Aust N Z J Med, 2000 Oct, 30(5), 578 - 84 Non-urease producing Helicobacter pylori in chronic gastritis; Ren Z et al.; BACKGROUND: Helicobacter pylori infection is the commonest cause of gastritis . Different patterns of immune response to H . pylori infection and characteristics of bacteria are considered to contribute to clinical outcomes . AIM: To determine characteristics of the host H . pylori relationship in subjects with non-ulcer dyspepsia and a histological diagnosis of gastritis . METHODS: Thirty-five subjects with chronic gastritis undergoing endoscopy (mean age 53 years, range 24-82, 14 male and 21 female) were studied, none of whom was on nonsteroidal anti-inflammatory drugs or antibiotics . H . pylori infection was determined by rapid urease test (CLOtest), culture, antibody and RT-PCR for Ure C, Cag A and 26 kDa gene and histology . Cytokine production of mucosal IL-6 and IL-8 were measured by ELISA . RESULTS: Fifteen subjects were positive by CLOtest and/or bacterial culture . In these subjects histology showed numerous helical forms of H . pylori (Group I) . Nine subjects were negative by CLOtest, bacterial culture, and mRNA for urease C fragment, but positive by PCR for the 26 kDa protein encoding gene . Histology in these subjects showed the presence of either coccoid forms (four), or scant helical forms (two), or mixed coccoid/helical forms (three) (Group II) . Eleven subjects were negative by all methods of detection (Group III) . IgG and IgA antibody levels in serum (p<0.05) and gastric tissue culture supernatant (p<0.001) were significantly higher in Group I than those in Group II or III . There were significant differences in the IgG serum and IgA supernatant antibody levels (p<0.01 and p<0.05) when Group II was compared to Group III . Supernatant IL-6 levels were significantly higher in Group I (p<0.01) than those from Groups II and III . IL-8 levels were higher in Group I (p<0.01) and Group II (p<0.05) when compared to Group III . CONCLUSIONS: 'H . pylori-negative' gastritis can be associated with a non-urease producing form of H . pylori, with a reduction in both local and systemic antibody levels and mucosal pro-inflammatory cytokines. J Electron Microsc (Tokyo), 2000, 49(2), 371 - 8 Reproducibility and applicability of gallium replication as evaluated by biological specimen use; Adachi E et al.; Structures of biological surfaces pressed on to a pure liquid gallium surface were successfully traced on to the gallium surface by quick-freezing below the melting point (28.78 degrees C) in air or water for replication in scanning electron microscopy . Gallium's high surface tension (approximately 700 mN m(-1) at 30 degrees C) deteriorates the spatial resolution of replicas and destroys some types of specimens . Five different biological surfaces were replicated on to gallium surfaces to evaluate spatial resolution and specimen resistance, i.e . reproducibility and applicability . Gallium replication of jewel beetle wing and human hair demonstrated submicron spatial resolution in the horizontal direction at least . Trials of protozoa, bacteria, and culture cell replication showed that protozoa are suited to replication because the cell membrane has characteristic structures with sufficient resistance to the gallium surface. Photochem Photobiol, 2000 Nov, 72(5), 639 - 44 Probing the primary event in the photocycle of photoactive yellow protein using photochemical hole-burning technique; Masciangioli T et al.; Photochemical hole-burning spectroscopy was used to study the excited-state electronic structure of the 4-hydroxycinnamyl chromophore in photoactive yellow protein (PYP) . This system is known to undergo a trans-to-cis isomerization process on a femtosecond-to-picosecond time scale, similar to membrane-bound rhodopsins, and is characterized by a broad featureless absorbance at 446 nm . Resolved vibronic structure was observed for the hole-burned spectra obtained when PYP in phosphate buffer at pH 7 was frozen at low temperature and irradiated with narrow bandwidth laser light at 431 nm . The approximate homogeneous width of 752 cm-1 could be calculated from the deconvolution of the hole-burned spectra leading to an estimated dephasing time of approximately 14 fs for the PYP excited-state structure . The resolved vibronic structure also enabled us to obtain an estimated change in the C=C stretching frequency, from 1663 cm-1 in the ground state to approximately 1429 cm-1 upon photoexcitation . The results obtained allowed us to speculate about the excited-state structure of PYP . We discuss the data for PYP in relation to the excited-state model proposed for the photosynthetic membrane protein bacteriorhodopsin, and use it to explain the primary event in the function of photoactive biological protein systems . Photoexcitation was also carried out at 475 nm . The vibronic structure obtained was quite different both in terms of the frequencies and Franck-Condon envelope . The origin of this spectrum was tentatively assigned. Med Dosw Mikrobiol, 2000, 52(1), 67 - 74 {Use of PCR methods for detection of selected genes of Mycoplasma pneumoniae}; Rastawicki W et al.; The aim of this study was standardization of PCR for the detection of gene encoding the P1 protein, 16S rRNA and elongation factor Tu of M . pneumoniae . A total of 13 strains of M . pneumoniae, 28 strains of other mycoplasmas and 14 strains of different bacteria causing respiratory tract infections were tested . In all of tested M . pneumoniae strains the presence of the sought genes was confirmed . The specificity of DNA was confirmed by the restriction endonuclease analysis with enzymes Hind III, Alu I and Hha I . With none of primers specific for the M . pneumoniae genes amplification of DNA from other bacteria was noted . The PCR method with the selected primers allowed to detect from 10(2) to 10(4) cfu M . pneumoniae/ml suspended in broth . The obtained results indicate that the PCR method can be used for detection of M . pneumoniae genes . A very good sensitivity and specificity predestine++ PCR as a potential quick diagnostic method for identification of M . pneumoniae in clinical specimens. Biochim Biophys Acta, 2000 Nov 20, 1460(2-3), 353 - 74 Interactions between redox partners in various cytochrome P450 systems: functional and structural aspects; Lewis DF et al.; The various types of redox partner interactions employed in cytochrome P450 systems are described . The similarities and differences between the redox components in the major categories of P450 systems present in bacteria, mitochondria and microsomes are discussed in the light of the accumulated evidence from X-ray crystallographic and NMR spectroscopic determinations . Molecular modeling of the interactions between the redox components in various P450 mono-oxygenase systems is proposed on the basis of structural and mutagenesis information, together with experimental findings based on chemical modification of key residues likely to be associated with complementary binding sites on certain typical P450 isoforms and their respective redox partners. Microbiol Mol Biol Rev, 2000 Dec, 64(4), 786 - 820 Origin and evolution of the mitochondrial proteome; Kurland CG et al.; The endosymbiotic theory for the origin of mitochondria requires substantial modification . The three identifiable ancestral sources to the proteome of mitochondria are proteins descended from the ancestral alpha-proteobacteria symbiont, proteins with no homology to bacterial orthologs, and diverse proteins with bacterial affinities not derived from alpha-proteobacteria . Random mutations in the form of deletions large and small seem to have eliminated nonessential genes from the endosymbiont-mitochondrial genome lineages . This process, together with the transfer of genes from the endosymbiont-mitochondrial genome to nuclei, has led to a marked reduction in the size of mitochondrial genomes . All proteins of bacterial descent that are encoded by nuclear genes were probably transferred by the same mechanism, involving the disintegration of mitochondria or bacteria by the intracellular membranous vacuoles of cells to release nucleic acid fragments that transform the nuclear genome . This ongoing process has intermittently introduced bacterial genes to nuclear genomes . The genomes of the last common ancestor of all organisms, in particular of mitochondria, encoded cytochrome oxidase homologues . There are no phylogenetic indications either in the mitochondrial proteome or in the nuclear genomes that the initial or subsequent function of the ancestor to the mitochondria was anaerobic . In contrast, there are indications that relatively advanced eukaryotes adapted to anaerobiosis by dismantling their mitochondria and refitting them as hydrogenosomes . Accordingly, a continuous history of aerobic respiration seems to have been the fate of most mitochondrial lineages . The initial phases of this history may have involved aerobic respiration by the symbiont functioning as a scavenger of toxic oxygen . The transition to mitochondria capable of active ATP export to the host cell seems to have required recruitment of eukaryotic ATP transport proteins from the nucleus . The identity of the ancestral host of the alpha-proteobacterial endosymbiont is unclear, but there is no indication that it was an autotroph . There are no indications of a specific alpha-proteobacterial origin to genes for glycolysis . In the absence of data to the contrary, it is assumed that the ancestral host cell was a heterotroph. Biochem J, 2000 Dec 15, 352 Pt 3, 659 - 66 Characterization of the unique function of a reduced amide bond in a cytolytic peptide that acts on phospholipid membranes; Oh JE et al.; The incorporation of a reduced amide bond, psi(CH(2)NH), into peptide results in an increase in the net positive charge and the perturbation of alpha-helical structure . By using this characteristic of the reduced amide bond, we designed and synthesized novel pseudopeptides containing reduced amide bonds, which had a great selectivity between bacterial and mammalian cells . A structure-activity relationship study on pseudopeptides indicated that the decrease in alpha-helicity and the increase in net positive charge in the backbone, caused by the incorporation of a reduced amide bond into the peptide, both contributed to an improvement in the selectivity between lipid membranes with various surface charges . However, activity results in vitro indicated that a perturbation of alpha-helical structure rather than an increase in net positive charge in the backbone is more important in the selectivity between bacterial and mammalian cells . The present result revealed that the backbone of membrane-active peptides were important not only in maintaining the secondary structure for the interactions with lipid membranes but also in direct interactions with lipid membranes . The present study showed the unique function of a reduced amide bond in cytolytic peptides and a direction for developing novel anti-bacterial agents from cytolytic peptides that act on the lipid membrane of micro-organisms. J Surg Res, 2000 Dec, 94(2), 178 - 84 Surgical stress induces phospholipid degradation in the intestinal brush border membrane; Prabhu R et al.; BACKGROUND: Surgical stress can lead to translocation of bacteria from the intestine into the systemic circulation . The intestinal brush border membrane (BBM) plays an important role in defense against such invasion by luminal bacteria and endotoxin . Our earlier work has shown the development of oxidative stress in the intestine after surgical stress and since the BBM is sensitive to free radical attack, this study examined the effect of surgical stress on the structure and function of intestinal BBM . METHODS: Intestinal BBM were isolated from control and after surgical stress and compared for structural and functional alterations . Surgical stress was also carried out following pretreatment with the xanthine oxidase inhibitor allopurinol or the nitric oxide donor l-arginine, to study the protection offered by these compounds . RESULTS: Surgical stress affected intestinal BBM structure as well as function . A decrease in alkaline phosphatase activity and alpha-tocopherol content, accompanied by an increase in lipid peroxidation, was seen . Surgical stress induced phospholipid degradation with generation of arachidonic acid . Functional impairment with a decrease in glucose transport ability was also seen . These changes are prevented by inhibition of xanthine oxidase by allopurinol pretreatment but not by NO . CONCLUSION: Surgical stress in the small intestine causes structural and functional alterations in the BBM through oxidative stress . This damage could affect gut barrier integrity and generation of arachidonic acid might mediate distal organ dysfunction . Braz J Infect Dis, 1997 Dec, 1(6), 306 - 313 Molecular Epidemiology of Lyme Disease Spirochetes Based on a Probe Complementary to Ribosomal RNA; dos Santos MA et al.; Lyme disease is caused by the spirochete, Borrelia burgdorferi, a bacteria which infects many vertebrates including humans . Borrelia have been isolated from many parts of the world, and there is interest to identify common genetic markers to improve molecular methods of diagnosis, and to aid in understanding varied manifestations of the disease . A total of 48 Borrelia burgdorferi strains, including: 38 isolated from ticks (Ixodes dammini, I . persulcatus, I . ricinus and I . pacificus), 3 from animals (dog, bird and hamster), and 7 from human clinical cases (skin, CSF, plasma and blood) from different geographic areas, were studied by DNA/DNA hybridization and rRNA gene restriction patterns by using a biotinylated pKK3535 probe (Altewegg M., Mayer L.W., 1989) . The migration patterns of rRNA gene-restriction fragments after clevage by Hind III separate these strains into 5 ribotypes of Borrelia burgdorferi: Type I (38 American,2 European strains); Type II (13 American strains); Type III (3 Asian and 1 European strains); Type IV (1 European and 2 Asian strains) and Type V (1 Asian strain) . The use of ribotyping has provided an additional tool to investigate the differences or common patterns which cause various Lyme disease syndromes. J Dairy Sci, 2000 Nov, 83(11), 2585 - 95 Nitrogen metabolism of early lactation cows fed diets with two different levels of protein and different amino acid profiles; Bach A et al.; Four multiparous Holstein cows (569+/-122 kg) surgically prepared with indwelling catheters in the mesenteric, portal, and hepatic veins and carotid artery were allocated in a 4 x 4 Latin square to determine the effects of dietary crude protein (CP) level and amino acid (AA) profile on N metabolism during early lactation (from 25 to 65 d in milk) . Cows received their diets in two equal meals and were milked twice daily . The dietary treatments were: 18% CP with a high (18H) or a low (18L) quality AA profile, and 15% CP with a high (15H) or a low (15L) quality AA profile . The four diets were similar in net energy for lactation (1.75 NEL Mcal/kg) and contained the same amount of RUP (34% of CP) . The quality of the AA profile pertained only to the essential AA (EAA), and was assessed by comparison with the EAA profile of casein and considered the potential contribution of EAA from ruminal bacteria . The 18H and 15H diets were supplemented with 50 and 25 g/d of ruminally protected Met, respectively . After 10 d on treatment, a blood flow marker (p-amino-hippurate) was infused into a mesenteric vein, and arterial, portal, hepatic, and mammary blood samples were obtained at 3, 6, and 12 h after feeding . Dry matter intake was similar across treatments (23.4+/-0.5 kg/d) . Amino acid oxidation, and consequent urea production, in the liver were numerically greater with the 18% CP rations, and, as a result, arterial urea concentrations were greatest (P < 0.01) with these rations . The amount of total AA extracted by the mammary gland tended to be greater with the H than with the L diets (21.4 vs . 18.2 mmol/ h, respectively) . Milk yield tended to be greater (P = 0.16) with the 18H and 15H diets (47.7 and 46.3 kg/d, respectively) compared with the 18L and 15L diets (45.9 and 44.6 kg/d, respectively) . Also, milk CP and casein contents were greatest (P = 0.09) with the H diets compared with the L diets . Milk and plasma urea N were greatest (P < 0.01) with the 18% CP diets . The efficiency of N utilization for milk protein synthesis was greatest (P < 0.09) with the 15% CP diets . It is concluded that milk protein production during early lactation is less susceptible to variations in dietary CP contents than variations in the AA profile of the dietary protein. Acta Cardiol, 2000 Oct, 55(5), 295 - 300 Chlamydia and atherosclerotic coronary arterial disease in Turkey; Ozsan M et al.; OBJECTIVE: Chlamydia pneumoniae, which is a Gram(-) intracellular bacteria, besides being a respiratory pathogen, is thought to play an active role in the progress of acute myocardial infarction and chronic coronary artery disease . In this study we aim to determine the frequency of C . pneumoniae in coronary artery lesions of Turkish people . METHODS AND RESULTS: The atherosclerotic material taken from 8 cases by directional atherectomy and from 23 cases by surgical endarterectomy and examined by indirect immunofluorescence (IIFA) test and polymerase chain reaction (PCR) . C . pneumoniae positivity was 32.3% (10/31) by IIFA and 29.0% (9/31) by PCR while the evaluation of the methods together yield a positivity of 35.5% (11/31) . CONCLUSIONS: A statistically significant difference could not be established between C . pneumoniae positive and negative groups according to age and the classical atherosclerotic risk factors such as diabetes mellitus, smoking, hypercholesterolaemia, hypertension, family history; besides, a statistically significant difference could not be found between the presence of C . pneumoniae and the severity and clinical picture of coronary artery disease. Curr Opin Biotechnol, 2000 Dec, 11(6), 540 - 6 Biodesulfurization and the upgrading of petroleum distillates; Monticello DJ; Biotechnology offers an alternative way to process fossil fuels . There have been several important advances in the elucidation of the mechanisms of biodesulfurization and the development of a biocatalytic desulfurization process . These include a detailed analysis of the rate and extent of desulfurization of real target molecules in a diesel matrix, the directed evolution of rate- and extent-limiting enzymes for better performance and the expression of the genes in alternative hosts . Process innovations include new reactor designs, separations and recovery strategies and the production of value-added byproducts during desulfurization. Curr Opin Immunol, 2000 Dec, 12(6), 632 - 40 The influence of infections on the development and severity of allergic disorders; Herz U et al.; The frequency and severity of atopic disorders are steadily increasing, particularly in developing countries . The reason for this observation is not clear . Recent studies indicate that infections with viruses and especially with bacteria early in life may help to inhibit allergic Th2 responses by skewing the immune system towards Th1 responses . However, infections can also lead to the exacerbation of atopic disorders. Nat Biotechnol, 2000 Dec, 18(12), 1311 - 4 Stable correction of a genetic deficiency in human cells by an episome carrying a 115 kb genomic transgene; Wade-Martins R et al.; Persistent expression of a transgene at therapeutic levels is required for successful gene therapy, but many small vectors with heterologous promoters are prone to vector loss and transcriptional silencing . The delivery of genomic DNA would enable genes to be transferred as complete loci, including regulatory sequences, introns, and native promoter elements . These elements may be critical to ensure prolonged, regulated, and tissue-specific transgene expression . Many studies point to considerable advantages to be gained by using complete genomic loci in gene expression . Large-insert vectors incorporating elements of the bacterial artificial chromosome (BAC) cloning system, and the episomal maintenance mechanisms of Epstein-Barr virus (EBV), can shuttle between bacteria and mammalian cells, allowing large genomic loci to be manipulated conveniently . We now demonstrate the potential utility of such vectors by stably correcting a human genetic deficiency in vitro . When the complete hypoxanthine phosphoribosyltransferase (HPRT) locus of 115 kilobases (kb) was introduced into deficient human cells, the transgene was both maintained as an episome and expressed stably for six months in rapidly dividing cell cultures . The results demonstrate for the first time that gene expression from an episomal genomic transgene can correct a cell culture disease phenotype for a prolonged period. J Clin Microbiol, 2000 Dec, 38(12), 4665 - 7 Comparison of iodophor and alcohol pledgets with the Medi-Flex blood culture prep kit II for preventing contamination of blood cultures; Wilson ML et al.; Iodophor and alcohol pledgets were compared with the Medi-Flex Prep Kit II for skin disinfection before venipuncture . Of 12,367 blood cultures collected, 6,362 were done with conventional pledgets and 6, 005 were done with Medi-Flex kits . Contamination occurred in 351 of 6,362 blood cultures (5.5%; range, 3.7 to 8.1%) with conventional pledgets versus 328 of 6,005 (5.5%; range, 3.5 to 7.5%) with Medi-Flex kits. J Biomol NMR, 2000 Oct, 18(2), 83 - 100 Lipari-Szabo mapping: A graphical approach to Lipari-Szabo analysis of NMR relaxation data using reduced spectral density mapping; Andrec M et al.; In this paper, we explore connections between the Lipari-Szabo formalism and reduced spectral density mapping, and show how spectral density estimates can be associated with Lipari-Szabo parameters via a simple geometric construction which we call Lipari-Szabo mapping . This relationship can be used to estimate Lipari-Szabo parameters from spectral density estimates without the need for nonlinear optimization, and to perform 'model selection' in a graphical manner . The Lipari-Szabo map also provides insight into the Lipari-Szabo model, and allows us to determine when a given set of experimental spectral densities are inconsistent with the Lipari-Szabo formalism . Practical applications of Lipari-Szabo mapping in conjunction with more traditional analysis methods are discussed. J Gastroenterol Hepatol, 2000 Oct, 15 Suppl, G73 - 7 Reprocessing of flexible endoscopes; Leung JW; Proper reprocessing of endoscopes prevents the risk of transmission of infection between patients . Meticulous mechanical cleaning is the most important step as it removes the majority of the contaminating bacteria . It should be performed before manual or automatic disinfection . High-level disinfection involves total immersion of the endoscope in a liquid chemical germicide (LCG) at a preset temperature and concentration for a pre-determined period of time . Subsequent rinsing and drying are essential steps to remove the chemical solution and prevent bacterial colonization during storage . Endoscopy units that are used for more than 50 procedures per week may benefit from cleaning in an automatic endoscope reprocessor (AER) . This allows automated exposure of the endoscope to the LCG with subsequent flushing and drying of the channels, and minimizes staff exposure to the LCG . Reprocessing should be performed by trained and accredited personnel according to written guidelines or standards of practice as defined by professional societies . Regular monitoring of the reprocessing process is important for quality control and in ensuring patients' safety. J Antibiot (Tokyo), 2000 Sep, 53(9), 879 - 85 Tubulysins, new cytostatic peptides from myxobacteria acting on microtubuli . Production, isolation, physico-chemical and biological properties; Sasse F et al.; New cytostatic compounds, tubulysins, were isolated from the culture broth of strains of the myxobacteria Archangium gephyra and Angiococcus disciformis . The compounds are peptides partly consisting of unusual amino acids and are distantly related to the dolastatins . The tubulysins were not active against bacteria and only little against fungi, but showed high cytostatic activity against mammalian cell lines with IC50 values in the picomolar range . An incubation with 50 ng/ml tubulysin A led to a complete disappearance of the microtubuli network of the cells within 24 hours . The more active tubulysin D induced multipolar spindles: At 0.5 ng/ml all mitotic cells showed more than four spindle poles. Scand J Gastroenterol, 2000 Oct, 35(10), 1048 - 52 Fructose- and sorbitol-reduced diet improves mood and gastrointestinal disturbances in fructose malabsorbers; Ledochowski M et al.; BACKGROUND: Fructose malabsorption is characterized by the inability to absorb fructose efficiently . As a consequence fructose reaches the colon where it is broken down by bacteria to short fatty acids, CO2 and H2 . Bloating, cramps, osmotic diarrhea and other symptoms of irritable bowel syndrome are the consequences and can be seen in about 50% of fructose malabsorbers . We have previously shown that fructose malabsorption is associated with early signs of mental depression and low serum tryptophan concentrations . It was therefore of interest whether a fructose-reduced diet could not only improve gastrointestinal complaints but also depressive signs seen in fructose malabsorbers . METHODS: Fifty-three adults (12 males, 41 females), who were identified as fructose malabsorbers according to their breath-H2 concentrations, filled out a Beck's depression inventory-questionnaire, and a questionnaire with arbitrary scales for measurement of meteorism, stool frequency and quality of life for a 4-week period before dietary intervention and 4 weeks after dietary change as for fructose- and sorbitol-reduced diet . RESULTS: Depression scores were reduced by 65.2% after 4 weeks of diet (P < 0.0001), and there was a significant reduction of meteorism (P < 0.0001) and stool frequency (P < 0.01) . Improvement of signs of depression and of meteorism was more pronounced in females than in males . CONCLUSION: Fructose- and sorbitol-reduced diet in subjects with fructose malabsorption does not only reduce gastrointestinal symptoms but also improves mood and early signs of depression. Nature, 2000 Nov 16, 408(6810), 325 - 30 Functional genomic analysis of C . elegans chromosome I by systematic RNA interference; Fraser AG et al.; Complete genomic sequence is known for two multicellular eukaryotes, the nematode Caenorhabditis elegans and the fruit fly Drosophila melanogaster, and it will soon be known for humans . However, biological function has been assigned to only a small proportion of the predicted genes in any animal . Here we have used RNA-mediated interference (RNAi) to target nearly 90% of predicted genes on C . elegans chromosome I by feeding worms with bacteria that express double-stranded RNA . We have assigned function to 13.9% of the genes analysed, increasing the number of sequenced genes with known phenotypes on chromosome I from 70 to 378 . Although most genes with sterile or embryonic lethal RNAi phenotypes are involved in basal cell metabolism, many genes giving post-embryonic phenotypes have conserved sequences but unknown function . In addition, conserved genes are significantly more likely to have an RNAi phenotype than are genes with no conservation . We have constructed a reusable library of bacterial clones that will permit unlimited RNAi screens in the future; this should help develop a more complete view of the relationships between the genome, gene function and the environment. J Microw Power Electromagn Energy, 2000, 35(3), 179 - 84 Complex high-frequency technology for protection of grain against pests; Mishenko AA et al.; The results of experimental investigation of physical methods are presented for suppressing of biological activity of grain and grain product pests: harmful insects at each developmental stage except eggs (Insecta), mites (Arachnida, Acariformes) and microscopic fungi and bacteria . The technologies under development for disinfestation and disinfection of grain are based on irradiation of grain by modulated pulses of high-frequency (HF) electromagnetic fields and on simultaneous action of a complex of factors: vacuum and HF-field induced plasma . The threshold value of the electric field intensity for total insect mortality was found to be E = 4.0-5.0 kV/cm in the pulse mode at the base frequency of 47.5 MHz . When the combined technology is used, conditions are created in the irradiation chamber for HF-discharge and plasma formation, which are very strong factors influencing the biological organisms . These raise the energy (and cost) efficiency (approximately $2-3 per tonne of grain) of the combined technology for destruction of grain pests with complete environmental safety. DNA Cell Biol, 2000 Nov, 19(11), 689 - 96 Structure of the gene for uteroferrin; Vallet JL et al.; The published structure of the gene for uteroferrin differs from that of the human and mouse tartrate-resistant acid phosphatase (TRAP) genes . Polymerase chain reaction using genomic DNA as template and primers designed from exon 2 of the porcine uteroferrin gene amplified a product containing two previously undescribed introns . Because of these discrepancies, we cloned an EcoRI fragment from a porcine genomic BAC library containing the uteroferrin gene, and the region containing the uteroferrin gene was completely sequenced . The uteroferrin gene spanned 2.5 kb and contained five exons, which is similar to the structure previously reported for human and mouse TRAP genes but different from the published structure of the uteroferrin gene . Southern blotting of porcine genomic DNA digested with a variety of enzymes was consistent with the sequence that we obtained . The most likely explanation for the differing results is that the previously reported structure for the uteroferrin gene was the result of artifactual elimination of introns 2 and 3 by bacteria and artifactual recombination of the region upstream of the transcription start site of this gene. Appl Environ Microbiol, 2000 Dec, 66(12), 5353 - 9 A homologue of the tryptophan-rich sensory protein TspO and FixL regulate a novel nutrient deprivation-induced Sinorhizobium meliloti locus; Davey ME et al.; A nutrient deprivation-induced locus in Sinorhizobium meliloti strain 1021 was identified by use of a Tn5-luxAB reporter gene transposon . The tagged locus is comprised of two open reading frames (ORFs) designated ndiA and ndiB for nutrient deprivation-induced genes A and B . Comparison of the deduced amino acid sequences of both ndiA and ndiB to the protein databases failed to reveal similarity to any known genes . The expression of the ndi locus was found to be induced by carbon and nitrogen deprivation, osmotic stress, and oxygen limitation and during entry into stationary phase . To identify regulatory components involved in the control of ndi gene expression, a second round of mutagenesis was performed on the primary ndiB::Tn5-luxAB-tagged strain (C22) with transposon Tn1721 . A double-mutant strain was obtained that lacked ndi locus transcriptional activity under all of the inducing conditions tested . The Tn1721-tagged gene showed a high degree of similarity to tryptophan-rich sensory protein TspO from Rhodobacter sphaeroides, as well as to mitochondrial benzodiazepine receptor pK18 from mammals . Induction of the ndi::Tn5-luxAB reporter gene fusion was restored under all inducing conditions by introducing the tspO coding region, from either S . meliloti or R . sphaeroides, in trans . Furthermore, it was found that, in addition to tspO, fixL, which encodes the sensor protein of an oxygen-sensing two-component system, is required for full expression of the ndi locus, but only under low oxygen tension. Appl Environ Microbiol, 2000 Dec, 66(12), 5116 - 22 Community composition of marine bacterioplankton determined by 16S rRNA gene clone libraries and fluorescence in situ hybridization; Cottrell MT et al.; We determined the compositions of bacterioplankton communities in surface waters of coastal California using clone libraries of 16S rRNA genes and fluorescence in situ hybridization (FISH) in order to compare the community structures inferred from these two culture-independent approaches . The compositions of two clone libraries were quite similar to those of clone libraries of marine bacterioplankton examined by previous studies . Clones from gamma-proteobacteria comprised ca . 28% of the libraries, while approximately 55% of the clones came from alpha-proteobacteria, which dominated the clone libraries . The Cytophaga-Flavobacter group and three others each comprised 10% or fewer of the clone libraries . The community composition determined by FISH differed substantially from the composition implied by the clone libraries . The Cytophaga-Flavobacter group dominated 8 of the 11 communities assayed by FISH, including the two communities assayed using clone libraries . On average only 10% of DAPI (4', 6'-diamidino-2-phenylindole)-stained bacteria were detected by FISH with a probe for alpha-proteobacteria, but 30% of DAPI-stained bacteria appeared to be in the Cytophaga-Flavobacter group as determined by FISH . alpha-Proteobacteria were greatly overrepresented in clone libraries compared to their relative abundance determined by FISH, while the Cytophaga-Flavobacter group was underrepresented in clone libraries . Our data show that the Cytophaga-Flavobacter group can be a numerically dominant component of coastal marine bacterioplankton communities. Nucleosides Nucleotides Nucleic Acids, 2000 Aug, 19(8), 1249 - 64 The interaction of drugs with DNA gyrase: a model for the molecular basis of quinolone action; Heddle JG et al.; DNA gyrase supercoils DNA in bacteria . The fact that it is essential in all bacteria and absent from eukaryotes makes it an ideal drug target . We discuss the action of coumarin and quinolone drugs on gyrase . In the case of coumarins, the drugs are known to be competitive inhibitors of the gyrase ATPase reaction . From a combination of structural and biochemical studies, the molecular details of the gyrase-coumarin complex are well established . In the case of quinolones, the drugs are thought to act by stabilising a cleavage complex between gyrase and DNA that arrests polymerases in vivo . The exact nature of the gyrase-quinolone-DNA complex is not known; we propose a model for this complex based on structural and biochemical data. Clin Infect Dis, 2000 Dec, 31(6), 1343 - 8 Epub 2000 Nov 29. Endotoxin, cytokines, and procalcitonin in febrile patients admitted to the hospital: identification of subjects at high risk of mortality; van Langevelde P et al.; We prospectively examined 464 febrile patients (median age, 61 years) for predictors of in-hospital death, by use of univariate and multivariate logistic regression using clinical data (age, underlying disease, duration of fever, chills, and shock on admission) and plasma endotoxin, TNF-alpha, IL-6, IL-10, and procalcitonin levels . The mortality rate was 4.6-fold higher (95% confidence interval {CI}, 1.8-12) in 31 patients with shock on admission, 7 of whom died; the strongest association with mortality was the endotoxin concentration (relative risk, 13.7; 95% CI, 1 . 4-136), which predicted 5 of the deaths with a 5% false-positive rate . For 433 patients without shock on admission, mortality (26 deaths) was associated with age and underlying disease: clinical data predicted 30% of the deaths, whereas IL-6 and procalcitonin levels identified an extra 10% with a 5% false-positive rate . When febrile patients are screened on hospital admission to identify those with a high risk for mortality, clinical judgment on the basis of age, underlying disease, and recent history outweighs the predictive value of endotoxin, cytokine, and procalcitonin levels . Only in patients who present with shock will measurement of endotoxin levels help predict those who will likely die at the cost of few false-positive results. Am J Hum Genet, 2001 Jan, 68(1), 208 - 13 Epub 2000 Nov 28. A mutation in the gene for the neurotransmitter receptor-clustering protein gephyrin causes a novel form of molybdenum cofactor deficiency; Reiss J et al.; Gephyrin was originally identified as a membrane-associated protein that is essential for the postsynaptic localization of receptors for the neurotransmitters glycine and GABA(A) . A sequence comparison revealed homologies between gephyrin and proteins necessary for the biosynthesis of the universal molybdenum cofactor (MoCo) . Because gephyrin expression can rescue a MoCo-deficient mutation in bacteria, plants, and a murine cell line, it became clear that gephyrin also plays a role in MoCo biosynthesis . Human MoCo deficiency is a fatal disease resulting in severe neurological damage and death in early childhood . Most patients harbor MOCS1 mutations, which prohibit formation of a precursor, or carry MOCS2 mutations, which abrogate precursor conversion to molybdopterin . The present report describes the identification of a gephyrin gene (GEPH) deletion in a patient with symptoms typical of MoCo deficiency . Biochemical studies of the patient's fibroblasts demonstrate that gephyrin catalyzes the insertion of molybdenum into molybdopterin and suggest that this novel form of MoCo deficiency might be curable by molybdate supplementation. Proc Natl Acad Sci U S A, 2000 Dec 5, 97(25), 13561 - 6 Crystal structures of photosynthetic reaction center and high-potential iron-sulfur protein from Thermochromatium tepidum: thermostability and electron transfer; Nogi T et al.; The reaction center (RC) of photosynthetic bacteria is a membrane protein complex that promotes a light-induced charge separation during the primary process of photosynthesis . In the photosynthetic electron transfer chain, the soluble electron carrier proteins transport electrons to the RC and reduce the photo-oxidized special-pair of bacteriochlorophyll . The high-potential iron-sulfur protein (HiPIP) is known to serve as an electron donor to the RC in some species, where the c-type cytochrome subunit, the peripheral subunit of the RC, directly accepts electrons from the HiPIP . Here we report the crystal structures of the RC and the HiPIP from Thermochromatium (Tch.) tepidum, at 2.2-A and 1.5-A resolution, respectively . Tch . tepidum can grow at the highest temperature of all known purple bacteria, and the Tch . tepidum RC shows some degree of stability to high temperature . Comparison with the RCs of mesophiles, such as Blastochloris viridis, has shown that the Tch . tepidum RC possesses more Arg residues at the membrane surface, which might contribute to the stability of this membrane protein . The RC and the HiPIP both possess hydrophobic patches on their respective surfaces, and the HiPIP is expected to interact with the cytochrome subunit by hydrophobic interactions near the heme-1, the most distal heme to the special-pair. Nucleic Acids Res, 2000 Dec 1, 28(23), 4623 - 33 Divergent mechanisms of 5' 23S rRNA IVS processing in the alpha-proteobacteria; Zahn K et al.; Widespread occurrence of a separate small RNA derived from the 5'-end of 23S rRNA and of an intervening sequence (IVS) which separates this domain from the main segment of 23S rRNA in the alpha-proteobacteria implies that processing reactions which act to excise the IVS are also maintained in this group . We previously characterized the first example of processing of this IVS in Rhodopseudomonas palustris, which is classified with the Bradyrhizobia In this case, IVS excision occurs by a multistep process and RNase III appears to act at an early step . Here, we characterize in vivo and in vitro IVS processing in two other related, but phenotypically distinct, Bradyrhizobia We also examine in vivo and in vitro processing of rRNA precursors from a more distantly related alpha-proteobacterium, Rhodobacter sphaeroides which produces a separate 5' 23S rRNA domain but has different sequences in the 5' 23S rRNA IVS . The details of the in vivo processing of all of the Bradyrhizobial rRNAs closely resemble the R . palustris example and in vitro studies suggest that all of the Bradyrhizobia utilize RNase III in the first step of IVS cleavage . Remarkably, in vivo and in vitro studies with R.sphaeroides indicate that initial IVS cleavage uses a different mechanism . While the mechanism of IVS cleavage differs among these alpha-proteobacteria, in all of these cases the limits of the internal segments processed in vivo are almost identical and occur far beyond the initial cleavage sites within the IVSs . We propose that these bacteria possess common secondary maturation pathways which enable them to generate similarly processed 23S rRNA 5'- and 3'-ends. Cornea, 2000 Nov, 19(6), 864 - 9 Infiltrative keratitis associated with extended wear of hydrogel lenses and Abiotrophia defectiva; Keay L et al.; PURPOSE: Infiltrative keratitis is a common complication associated with extended wear of hydrogel lenses . Causative bacteria are often isolated from the lens at the time of an event . We report a case where three repeated occurrences of infiltrative keratitis were associated with contamination of the contact lenses by Abiotrophia defectiva . METHODS: A 34-year-old man participating in a clinical trial of extended wear hydrogel contact lenses experienced three episodes of infiltrative keratitis . The clinical presentation was observed using a biomicroscope . At the time of each event, the contact lenses were removed aseptically and ocular swabs were taken for bacterial identification and enumeration . The condition was monitored until full resolution . RESULTS: The condition was characterized by irritation, marked bulbar and limbal injection, and multiple focal subepithelial infiltrates . Many of the infiltrates also showed overlying staining with fluorescein . In each of the three events of infiltrative keratitis, A . defectiva was cultured from the contact lens and ocular swabs . CONCLUSION: This is the first reported occurrence of infiltrative keratitis associated with A . defectiva contamination of contact lenses. FEBS Lett, 2000 Nov 24, 485(2-3), 113 - 6 Iron starvation leads to increased expression of Cu/Zn-superoxide dismutase in Aspergillus; Oberegger H et al.; In a search for iron-regulated proteins of Aspergillus nidulans and Aspergillus fumigatus a 16-kDa protein was identified which is about 5-fold upregulated during iron starvation in both species and which can be approximately 500-fold enriched by simple one-step chromatography on Amberlite XAD-16 resin . N-terminal protein sequence analysis and cloning of the respective A . nidulans cDNA identified this protein as a Cu/Zn-superoxide dismutase (SODA) . Northern analysis revealed that upregulation of sodA expression occurs at the level of transcript accumulation . This seems to be a specific low iron response and not a general starvation answer since sodA transcript levels do not respond to carbon or nitrogen starvation . In contrast, copper depletion leads to transcriptional downregulation of sodA . Furthermore, sodA expression was found still to be subject to iron regulation in an A . nidulans mutant lacking SREA, a regulator of iron homeostasis, indicating that sodA expression is regulated by an SREA-independent mechanism . The data presented suggest that SODA plays a protective role under iron deplete conditions. J Biol Chem, 2001 Mar 16, 276(11), 7721 - 6 Epub 2000 Nov 28. Interaction between the N-terminal domain of gastric H,K-ATPase and the spectrin binding domain of ankyrin III; Festy F et al.; We screened a cDNA bank of rabbit gastric fundic mucosa by two-hybrid assays looking for binding partners of the N-terminal domain of the rabbit gastric H,K-ATPase . We extracted five clones sharing more than 90% sequence identity . The longest clone codes for a protein sharing a high identity (96 and 96.8%, respectively) with a fragment of the membrane domain, from Arg-835 to Ser-873, plus the major part of the "spectrin binding domain" going from Glu-874 to Leu-1455 of human and mouse ankyrin III . We conclude that the membrane and spectrin binding domains of the rabbit ankyrin III are candidates for the binding partner of the N-terminal domain of the rabbit gastric H,K-ATPase . To validate the ankyrin-ATPase interaction and to test its specificity, we produced both domains in yeast and bacteria, coimmunoprecipitated them with an anti-ATPase antibody, and copurified them by affinity chromatography . The sequence of rabbit ankyrin III was not known, and this is the first report demonstrating that the ankyrin III and the H,K-ATPase interact with no intermediate . The interaction involves the N-terminal domain of the ATPase on one hand and the spectrin binding domain of the ankyrin on the other. Carbohydr Res, 2000 Oct 6, 328(4), 605 - 10 Synthesis of furanose derivatives of 3-deoxy-D-erythro-2-hexulosonic acid and their 3-bromo and 3-deuterio analogs; Di Nardo C et al.; Methanolysis of 2,4,6-tri-O-benzoyl-2,3-dibromo-3-deoxy-D-altrono-1,5-lactone gave methyl 3-bromo-3-deoxy-2,4,6-tri-O-benzoyl-alpha-D-ribo-hex-2-ulofuranosonat e (3) and the anomeric mixture of the analogous 4,6-di-O-benzoyl derivative, having HO-2 free . Compound 3 was subjected to debromination with tributyltin hydride and tributyltin deuteride in the presence of 2,2'-azo-bisisobutyronitrile affording, respectively, the corresponding derivatives of 3-deoxy-D-erythro-2-hexulosonic acid and its 3-deuterio analog . The structure of the products and intermediates was established by spectroscopic methods and chemical transformations. Expert Opin Investig Drugs, 2000 Dec, 9(12), 2783 - 97 Antineoplastic agents from natural sources: achievements and future directions; Cragg GM et al.; The influence of natural products upon anticancer drug discovery and design cannot be overestimated . Approximately 60% of all drugs now in clinical trials for the multiplicity of cancers are either natural products, compounds derived from natural products, contain pharmacophores derived from active natural products or are 'old drugs in new clothes', where (modified) natural products are attached to targeting systems . This review covers those materials that the authors are aware of as being in clinical trials through early 2000 and demonstrates how, even today, in the presence of massive numbers of agents from combinatorial libraries, the compounds produced by 'Mother Nature' are still in the forefront of cancer chemotherapeutics as sources of active chemotypes. Jpn J Cancer Res, 2000 Nov, 91(11), 1161 - 8 Induction of apoptosis and cell cycle arrest in mouse colon 26 cells by benastatin A; Kakizaki I et al.; Benastatin A, isolated from Streptomyces bacteria, is reported to inhibit mammalian glutathione transferases (GSTs) . Since GST inhibitors such as ethacrynic acid are suggested to induce apoptosis in some cell lines, the effect of benastatin A on the survival of mouse colon 26 adenocarcinoma cells was compared with that of ethacrynic acid . When cells in stationary phase were treated with benastatin A, viable cells were found to be dose-dependently decreased after 3 days . In the case of ethacrynic acid, this became apparent within 24 h . Electrophoretic analysis revealed DNA fragmentation, indicating that cell loss was due to apoptosis in both cases . The dominant GST in colon 26 cells was identified as the class Pi-form (GST-II), and the activities in crude extracts as well as purified GST-II were almost completely inhibited by 50 microM ethacrynic acid . Immunoblot and northern blot analyses revealed increased GST-II protein and mRNA levels in cells treated with ethacrynic acid . Benastatin A did not significantly affect the activity in the crude extract even at 20 microM, a 10-fold higher concentration than that which almost completely inhibited the activity of purified GST-II . However, GST activity and GST-II protein were decreased in colon 26 cells treated with benastatin A for 5 days, no significant activity being detected in the range of 16 - 20 microM . In addition, beta-actin and bax mRNAs were also decreased in a dose-dependent manner . Furthermore, flow cytometric analysis of colon 26 cells revealed that benastatin A blocked the cell cycle at the G1/G0 phase . Thus, benastatin A also induces apoptosis of colon 26 cells, but this is unlikely to be due to inhibition of GST activity. Acta Crystallogr D Biol Crystallogr, 2000 Dec, 56 Pt 12, 1634 - 7 Crystallization of the Mycobacterium tuberculosis cell-division protein FtsZ; Leung AK et al.; Mycobacterium tuberculosis FtsZ (MtbFtsZ), an essential protein in bacterial cell division, has been crystallized in the presence of a new inhibitor of MtbFtsZ polymerization and GTPase activity, ethyl (6-amino-2,3-dihydro-4-phenyl-1H-pyrido{4,3-b}{1, 4}diazepin-8-yl)carbamate (SRI-7614) . Crystals of the MtbFtsZ-SRI-7614 complex (form I, 30% polyethylene glycol 4000, 0.1 M sodium citrate pH 5.6, 0.2 M NH(4)OAc, 293 K) belong to space group P6(1) or P6(5), with unit-cell parameters a = 88.78, c = 178 . 02 A, and diffract to 2.3 A resolution . A second crystal form, of the GDP complex, grows in the presence or absence of Mg(2+) from PEG 4000 at 277 K or from (NH(4))(2)SO(4) at 293 K, respectively (form II, space group P6(2)22 or P6(4)22, with unit-cell parameters a = 135.02, c = 328.97 A or a = 129.30, c = 327.97 A, respectively) . Complete data sets to approximately 7 A resolution have been collected from both . Exceptional form II crystals diffract to at least 4.5 A resolution . Determination of the MtbFtsZ structure may advance the design of improved inhibitors of FtsZ polymerization. Acta Crystallogr D Biol Crystallogr, 2000 Dec, 56 Pt 12, 1554 - 9 Ab initio phasing of a 4189-atom protein structure at 1.2 A resolution; Tame JR; The phase problem remains a key rate-limiting step in the determination of macromolecular X-ray structures . Direct methods, applying probability theory to the native data set, can routinely solve structures of up to about 200 non-H atoms, although much larger structures have been solved given sufficiently high resolution data and the presence of heavy atoms . Here it is shown that maximum-likelihood refinement of free-atom models with ARP/wARP can solve ab initio a much larger metalloprotein structure than the largest so far solved by conventional direct methods . The protein, OppA, is not naturally associated with metal ions but was co-crystallized with uranium. Annu Rev Genet, 2000, 34, 359 - 399 DNA mismatch repair and genetic instability; Harfe BD et al.; Mismatch repair (MMR) systems play a central role in promoting genetic stability by repairing DNA replication errors, inhibiting recombination between non-identical DNA sequences and participating in responses to DNA damage . The discovery of a link between human cancer and MMR defects has led to an explosion of research on eukaryotic MMR . The key proteins in MMR are highly conserved from bacteria to mammals, and this conservation has been critical for defining the components of eukaryotic MMR systems . In eukaryotes, there are multiple homologs of the key bacterial MutS and MutL MMR proteins, and these homologs form heterodimers that have discrete roles in MMR-related processes . This review describes the genetic and biochemical approaches used to study MMR, and summarizes the diverse roles that MMR proteins play in maintaining genetic stability. Angew Chem Int Ed Engl, 2000 Nov 17, 39(22), 4004 - 4032 Piezoelectric Mass-Sensing Devices as Biosensors-An Alternative to Optical Biosensors? Janshoff A, Galla HJ, Steinem C. In the early days of electronic communication-as a result of the limited number of quartz resonators available-frequency adjustment was accomplished by a pencil mark depositing a foreign mass layer on the crystal . In 1959, Sauerbrey showed that the shift in resonance frequency of thickness-shear-mode resonators is proportional to the deposited mass . This was the starting point for the development of a new generation of piezoelectric mass-sensitive devices . However, it was the development of new powerful oscillator circuits that were capable of operating thickness shear mode resonators in fluids that enabled this technique to be introduced into bioanalytic applications . In the last decade adsorption of biomolecules on functionalized surfaces turned in to one of the paramount applications of piezoelectric transducers . These applications include the study of the interaction of DNA and RNA with complementary strands, specific recognition of protein ligands by immobilized receptors, the detection of virus capsids, bacteria, mammalian cells, and last but not least the development of complete immunosensors . Piezoelectric transducers allow a label-free detection of molecules; they are more than mere mass sensors since the sensor response is also influenced by interfacial phenomena, viscoelastic properties of the adhered biomaterial, surface charges of adsorbed molecules, and surface roughness . These new insights have recently been used to investigate the adhesion of cells, liposomes, and proteins onto surfaces, thus allowing the determination of the morphological changes of cells as a response to pharmacological substances and changes in the water content of biopolymers without employing labor-intense techniques . However, the future will show whether the quartz-crystal microbalance will assert itself against established label-free sensor devices such as surface plasmon resonance spectroscopy and interferometry. Autoimmunity, 2000 Oct, 32(3), 153 - 60 Do immune complexes formed with autoantibodies have a role in the maintenance of immune homeostasis through interaction with FC receptors; Guarnotta G et al.; Natural autoantibodies play an important regulatory role in the maintenance of immune homeostasis . They act as a first line of defense against environmental pathogens like toxins, bacteria and erythrocytes . In humans they are mainly produced by CD5+ B cells that are under the control of a regulatory T cell population . Fc-gamma receptors are involved in antigen recognition and signal transduction and tuning, and some of the members of the FcR family have structural similarity to MHC molecules; they may interact with multiple Ig ligands and with non-Ig ligands . We discuss the interactions between immune-complexes formed with natural autoantibodies and Fc-gamma receptors and suggest that such interactions may affect self-recognition in the thymus and regulate immune homeostasis. Toxicology, 2000 Nov 2, 152(1-3), 47 - 52 (1-->3)-beta-D-glucan - relationship to indoor air-related symptoms, allergy and asthma; Rylander R et al.; (1-->3)-beta-D-glucan is a polyglucose structure in the cell wall of moulds, some bacteria and plants . Due to its unique (1-->3)-beta linkage it binds to specific receptors on phagocytosing cells and induces changes in their metabolism . Under realistic environmental concentrations, available data suggest that these changes express themselves as alterations of the defense mechanisms to other agents . Inhalation of (1-->3)-beta-D-glucan in humans causes symptoms from the upper respiratory tract and induction of cytokines in blood monocytes . (1-->3)-beta-D-glucan can be used as a marker of mould biomass in field studies . Relationships between the amount of (1-->3)-beta-D-glucan and the extent of symptoms as well as lung function changes and inflammatory markers have been described . In view of the mechanisms involved in the normal development of the immune system, children seem to be a particular group at risk due to (1-->3)-beta-D-glucan exposure. Vaccine, 2000 Nov 8, 19(6), 637 - 43 Investigation of cellular and humoral immune responses to whole cell and acellular pertussis vaccines; Canthaboo C et al.; New generation acellular pertussis vaccines were compared with the established whole cell pertussis vaccine for the induction of humoral and cellular immune-responses in mice . At the same time, the in vivo protective effect of these two types of vaccine was also compared in both intracerebral (ic) and aerosol challenge models . In general, whole cell vaccine induced lower antibody titres to pertussis toxin, filamentous haemagglutinin and pertactin than the acellular vaccine . Nitric oxide concentration in macrophage cultures was used as a marker for macrophage activation . The nitric oxide concentrations in the macrophage cultures from mice following immunisation with the whole cell vaccine were higher than those from mice immunised with the acellular vaccine, which indicated that the whole cell vaccine was more effective than the acellular vaccine in activating macrophages . This was associated with better protection in vivo after challenge . After ic challenge of mice following immunisation with whole cell or acellular vaccine, 90% of the whole cell vaccine group survived compared with 40% of the acellular vaccine group at the vaccine dose selected . Following aerosol challenge, mice in the whole cell vaccine group showed faster clearance of bacteria from the lungs than those in the acellular vaccine group . Our findings suggest that the different types of pertussis vaccines may achieve protection in different ways and that CMI may play an important role in eliminating bacteria which escape humoral defence mechanisms. Aquat Toxicol, 2001 Feb, 51(4), 405 - 17 Effects of Al(3+) ions and Cu(2+) ions on microcosms with three different biological complexities; Sugiura K; Cu(2+) ions or Al(3+) ions were added to microcosms containing three, four and eight species (S-3, S-4, S-8) at the beginning of the culture, or at the 15th day from the beginning of the culture to determine the production rates (P), the respiration rates (R), and P/R ratios . There was a balance between the oxygen production rates and the oxygen consumption rates in the microcosms (S-4) and (S-8), whereas there was no balance between two quantities in the microcosm (S-3) . There were no significant differences in the Cu(2+) and Al(3+) ion concentrations influencing the material cycles and the metabolic balance between the case when the number of species was four and the case when the number of species was eight . It was assumed that the effects concentrations of the chemical substances are not significantly related to the degree of diversity of the constituent species in the microcosms having a balance between production and consumption. J Virol, 2000 Dec, 74(24), 11548 - 56 Integrase-lexA fusion proteins incorporated into human immunodeficiency virus type 1 that contains a catalytically inactive integrase gene are functional to mediate integration; Holmes-Son ML et al.; Purified fusion proteins made up of a retroviral integrase and a sequence-specific DNA-binding protein have been tested in in vitro assays for their ability to direct integration into specific target sites . To determine whether these fusion proteins can be incorporated into human immunodeficiency virus type 1 (HIV-1) and are functional to mediate integration, we used an in trans approach to deliver various integrase-LexA proteins to an integrase-defective virus containing an integrase mutation at aspartate residue 64 . Integrase-LexA, integrase-LexA DNA-binding domain, or N- or C-terminally truncated integrase-LexA proteins were fused to the HIV-1 accessory protein, Vpr . Coexpression of the Vpr fusion proteins and an integrase-defective HIV-1 molecular clone by a producer cell line resulted in efficient incorporation of the fusion protein into the integrase-mutated virus . In addition, each of these viruses was infectious and capable of performing integration, as determined by two independent cellular assays that measure reporter gene expression . With the exception of the N-terminally truncated integrase fused to LexA, which was at about 1%, all of the fusion proteins restored integration to a similar level, at 17 to 24% of that of the wild-type virus . The low level observed with the N-terminally truncated integrase fused to LexA is consistent with previous results implying that the N terminus of integrase is involved in multiple steps of the retroviral life cycle . These data indicate that the integrase-fusion proteins retain catalytic function in the integrase-mutated viruses and demonstrate the feasibility of incorporating integrase fusion proteins into HIV-1 for the development of site-directed retroviral vectors. Eur J Cell Biol, 2000 Oct, 79(10), 735 - 49 Disruption of the actin filament network affects delivery of endocytic contents marker to phagosomes with early endosome characteristics: the case of phagosomes with pathogenic mycobacteria; Guerin I et al.; Phagosomes containing live virulent mycobacteria undergo fusion with early endosomes, but they are unable to mature normally . Accordingly, they do not fuse with lysosomes . Although M . avium-containing phagosomes retain fusion and intermingling characteristics of early endosomes indefinitely, fusions with early endosomes are increasingly restricted as bacteria multiply . In addition, when endocytic tracers, such as horseradish peroxidase (HRP), are added to M . avium-infected macrophages at 1 or up to 15 days after infection, an atypical time course of acquisition of the tracer by the phagosomes is observed, i.e., a 10 to 20 min lag, instead of immediate acquisition as is typical for early endosomes (and phagosomes with early endosome characteristics) . These events coincide with a marked disorganization of the actin filament network in M . avium-infected macrophages . In the present study, we have therefore addressed the following question: Do actin filaments play a role in fusion and intermingling of contents between early endosomes and immature phagosomes that undergo homotypic fusion with early endosomes? We examined the time course of acquisition of subsequently internalized endocytic marker (HRP) by early endosome-like preexisting phagosomes, i.e . 2 hour-old phagosomes with either hydrophobic latex particles, virulent or avirulent M . avium, after depolymerization of the actin filament network with cytochalasin D or after repolymerization of the actin filament network with jasplakinolide, in cases where the network had been depolymerized (macrophages infected with M . avium, at 1 or up to 7 days after infection) . By direct morphological observation at the electron microscope level and by a kinetic approach, we show here that depolymerization of the actin filament network with cytochalasin D delays acquisition of HRP whereas repolymerization restores immediate acquisition of the marker . We conclude that the actin filament network is involved in fusion and intermingling of endocytic contents between early endosomes and early endosome-like phagosomes, and that disruption of this network by M . avium is the cause for the atypical acquisition of content marker by phagosomes containing these pathogenic mycobacteria. Phys Rev E Stat Phys Plasmas Fluids Relat Interdiscip Topics, 2000 Oct, 62(4 Pt B), 5353 - 9 Model for water transport into powdered xanthan combining gel swelling and vapor diffusion; Goerke U et al.; Water ingress into xanthan powder compressed to various packing densities has been studied using nuclear magnetic resonance stray field imaging (STRAFI) . A foot is observed ahead of the main water ingress front which is attributed to vapor transport around the particles . The main development of the reported work is an analytical model which describes the coupling of vapor transport through the pore space and liquid transport through the progressively swelling gel which gradually occludes the vapor path . Good agreement between theory and experiment is found over a wide range of packing densities with the model requiring only one adjustable parameter, the water diffusivity in the gel measured in a constant polymer mass reference frame . It is suggested that the results are of considerable relevance to situations where the polymer is produced at low concentration by bacteria such as in the rhizosphere and aerial bio films. Protein Expr Purif, 2000 Dec, 20(3), 421 - 34 Synthesis of a highly substituted N(6)-linked immobilized NAD(+) derivative using a rapid solid-phase modular approach: suitability for use with the kinetic locking-on tactic for bioaffinity purification of NAD(+)-dependent dehydrogenases; Tynan J et al.; This study is concerned with further development of the kinetic locking-on strategy for bioaffinity purification of NAD(+)-dependent dehydrogenases . Specifically, the synthesis of highly substituted N(6)-linked immobilized NAD(+) derivatives is described using a rapid solid-phase modular approach . Other modifications of the N(6)-linked immobilized NAD(+) derivative include substitution of the hydrophobic diaminohexane spacer arm with polar spacer arms (9 and 19.5 A) in an attempt to minimize nonbiospecific interactions . Analysis of the N(6)-linked NAD(+) derivatives confirm (i) retention of cofactor activity upon immobilization (up to 97%); (ii) high total substitution levels and high percentage accessibility levels when compared to S(6)-linked immobilized NAD(+) derivatives (also synthesized with polar spacer arms); (iii) short production times when compared to the preassembly approach to synthesis . Model locking-on bioaffinity chromatographic studies were carried out with bovine heart l-lactate dehydrogenase (l-LDH, EC 1.1.1.27), bakers yeast alcohol dehydrogenase (YADH, EC 1.1.1.1) and Sporosarcinia sp . l-phenylalanine dehydrogenase (l-PheDH, EC 1.4.1.20), using oxalate, hydroxylamine, and d-phenylalanine, respectively, as locking-on ligands . Surprisingly, two of these test NAD(+)-dependent dehydrogenases (lactate and alcohol dehydrogenase) were found to have a greater affinity for the more lowly substituted S(6)-linked immobilized cofactor derivatives than for the new N(6)-linked derivatives . In contrast, the NAD(+)-dependent phenylalanine dehydrogenase showed no affinity for the S(6)-linked immobilized NAD(+) derivative, but was locked-on strongly to the N(6)-linked immobilized derivative . That this locking-on is biospecific is confirmed by the observation that the enzyme failed to lock-on to an analogous N(6)-linked immobilized NADP(+) derivative in the presence of d-phenylalanine . This differential locking-on of NAD(+)-dependent dehydrogenases to N(6)-linked and S(6)-linked immobilized NAD(+) derivatives cannot be explained in terms of final accessible substitutions levels, but suggests fundamental differences in affinity of the three test enzymes for NAD(+) immobilized via N(6)-linkage as compared to thiol-linkage . J Nat Prod, 2000 Nov, 63(11), 1570 - 2 New butenolides from two marine streptomycetes; Mukku VJ et al.; Chemical examination of two marine Streptomycetes has resulted in the isolation of four new butenolides, namely 4, 10-dihydroxy-10-methyl-dodec-2-en-1,4-olide (1), two diastereomeric 4,11-dihydroxy-10-methyl-dodec-2-en-1,4-olides (2/3), and 4-hydroxy-10-methyl-11-oxo-dodec-2-en-1,4-olide (4) . The structures were identified by interpretation of the 2D NMR and mass spectral data. Ann Med, 2000 Oct, 32(7), 475 - 84 Efforts in minimizing risk of viral transmission through viral inactivation; Horowitz B et al.; The viral safety of blood and blood products has improved substantially over the last decade on account of the development of new viral screening and virucidal procedures . For nearly 15 years, virally inactivated blood derivatives, prepared by using advanced virucidal procedures, have amassed an extraordinary safety record with respect to hepatitis B and C and HIV . This record of safety has spawned the development of newer virucidal procedures designed to eliminate nonenveloped viruses from blood derivatives and viruses and other pathogens from blood components, including cellular components . Ongoing tests that include clinical studies will demonstrate how close we are to achieving a blood supply that is free of viruses, bacteria, and parasites. Vet Q, 2000 Oct, 22(4), 204 - 8 Defence mechanisms against viral infection in poultry: a review; Jeurissen SH et al.; Defence against viral infections in poultry consists of innate and adaptive mechanisms . The innate defence is mainly formed by natural killer cells, granulocytes, and macrophages and their secreted products, such as nitric oxide and various cytokines . The innate defence is of crucial importance early in viral infections . Natural killer cell activity can be routinely determined in chickens of 4 weeks and older using the RP9 tumour cell line . In vitro assays to determine the phagocytosis and killing activity of granulocytes and macrophages towards bacteria have been developed for chickens, but they have not been used with respect to virally infected animals . Cytokines, such as interleukin (IL)-1, IL-6 and tumour necrosis factor (TNF)-alpha, are indicators of macrophage activity during viral infections, and assays to measure IL-1 and IL-6 have been applied to chicken-derived materials . The adaptive defence can be divided into humoral and cellular immunity and both take time to develop and thus are more important later on during viral infections . Various enzyme-linked immunosorbent assays (ELISAs) to measure humoral immunity specific for the viruses that most commonly infect poultry in the field are now commercially available . These ELISAs are based on a coating of a certain virus on the plate . After incubation with chicken sera, the bound virus-specific antibodies are recognized by conjugates specific for chicken IgM and IgG . Cytotoxic T lymphocyte activity can be measured using a recently developed in vitro assay based on reticuloendotheliosis virus-transformed target cells that are loaded with viral antigens, e.g . Newcastle disease virus . This assay is still in an experimental stage, but will offer great opportunities in the near future for research into the cellular defence mechanisms during viral infections. J Immunol, 2000 Dec 1, 165(11), 6148 - 55 Vaccination with empty plasmid DNA or CpG oligonucleotide inhibits diabetes in nonobese diabetic mice: modulation of spontaneous 60-kDa heat shock protein autoimmunity; Quintana FJ et al.; Nonobese diabetic (NOD) mice develop insulitis and diabetes through a process involving autoimmunity to the 60-kDa heat shock protein (HSP60) . Treatment of NOD mice with HSP60 or with peptides derived from HSP60 inhibits this diabetogenic process . We now report that NOD diabetes can be inhibited by vaccination with a DNA construct encoding human HSP60, with the pcDNA3 empty vector, or with an oligonucleotide containing the CpG motif . Prevention of diabetes was associated with a decrease in the degree of insulitis and with down-regulation of spontaneous proliferative T cell responses to HSP60 and its peptide p277 . Moreover, both the pcDNA3 vector and the CpG oligonucleotide induced specific Abs, primarily of the IgG2b isotype, to HSP60 and p277, and not to other islet Ags (glutamic acid decarboxylase or insulin) or to an unrelated recombinant Ag expressed in bacteria (GST) . The IgG2b isotype of the specific Abs together with the decrease in T cell proliferative responses indicate a shift of the autoimmune process to a Th2 type in treated mice . These results suggest that immunostimulation by bacterial DNA motifs can modulate spontaneous HSP60 autoimmunity and inhibit NOD diabetes. Biochem J, 2000 Dec 1, 352 Pt 2, 343 - 51 Cloning and expression of a cDNA encoding human inositol 1,4,5-trisphosphate 3-kinase C; Dewaste V et al.; Inositol 1,4,5-trisphosphate {Ins(1,4,5)P(3)} 3-kinase catalyses the phosphorylation of Ins(1,4,5)P(3) to Ins(1,3,4,5)P(4) . cDNAs encoding two isoenzymes of Ins(1,4,5)P(3) 3-kinase (3-kinases A and B) have been described previously . In the present study, we report the cloning of a full-length 2052 bp cDNA encoding a third human isoenzyme of the Ins(1,4,5)P(3) 3-kinase family, referred to as isoform C . This novel enzyme has a calculated molecular mass of 75 . 207 kDa and a K(m) for Ins(1,4,5)P(3) of 6 microM . Northern-blot analysis showed the presence of a transcript of approx . 3.9 kb in various human tissues . Inositol trisphosphate 3-kinase C demonstrates enzymic activity when expressed in DH5alphaF' bacteria or COS-7 cells . Calcium alone decreases the Ins(1,4,5)P(3) 3-kinase activity of the 3-kinase C isoenzyme in transfected COS-7 cells . This inhibitory effect is reversed in the presence of calmodulin . The recombinant bacterial 3-kinase C can be adsorbed on calmodulin-Sepharose in the presence of calcium . The present data show that Ins(1,4,5)P(3) 3-kinase C: (i) shares a conserved catalytic domain of about 275 amino acids with the two other mammalian isoforms, (ii) could be purified on a calmodulin-Sepharose column and (iii) could be distinguished from the A and B isoenzymes by the effects of calcium and of calmodulin. J Submicrosc Cytol Pathol, 2000 Apr, 32(2), 159 - 67 Structure of lamina propria lymphoid follicles and associated epithelium in the gastric mucosa during Helicobacter pylori infection in ulcer-bearing Mongolian gerbils; Wada R et al.; To develop a gerbil model of Helicobacter pylori-induced chronic active gastritis comparable in severity to human lesions, we made acetic acid-induced ulcer in the anterior antral wall and concurrently challenged 1 x 10(8) colony-forming units bacteria per os . At 30 and 60 days after inoculation, the number of viable bacteria colonizing on the surface epithelium of the gastric mucosa was larger in ulcer-bearing animals compared to non-bearing ones . Furthermore, in the former animals, neutrophil and mononuclear cell infiltration as well as lymphoid follicle formation in the lamina propria was more prominent . Electron microscopically, lymphoid follicle-associated epithelium displayed specialized structures . Namely, brush cells interposed between mucous epithelial cells and characterized by prominent microfilament bundles and many apical vesicles or caveola specifically embraced the cluster of intraepithelially invading lymphocytes and macrophage-like cells by the attenuated cytoplasm in an analogous manner to M cells in Peyer's patches . The present study has demonstrated that ulcer formation enhances both H . pylori colonization and lamina propria lymphoid follicle formation and suggested that follicle-associated epithelium might play roles in the delivery of intraluminal antigen. Vet Clin North Am Food Anim Pract, 2000 Nov, 16(3), 497 - 509, vii Ionophore use and toxicosis in cattle; Hall JO; Ionophores comprise a rapidly expanding class of antibiotics produced by filamentous branching bacteria of the order Actinomycetales . The use of ionophores as coccidiostats and growth promotants has resulted in the occurrence of toxicoses in target and nontarget species . Clinical and pathologic effects of ionophore poisoning are caused by bioactivity and damage to excitable tissues such as cardiac muscle, skeletal muscle, smooth muscle, and the nervous system . Ionophore toxicoses are often related to errors in feed mixing, so the practitioner should give primary importance to the removal of suspect feeds and testing to confirm excessive exposure. Vet Clin North Am Food Anim Pract, 2000 Nov, 16(3), 455 - 64 Current knowledge of water quality and safety for livestock; Carson TL; Basic laboratory evaluation of water quality for livestock should include measurement of TDS, sulfate, nitrate-nitrite, and coliform bacteria . Supplementary water tests may include pH, sodium, iron, magnesium, chloride, calcium, potassium, manganese, and contaminants specific to the situation . Using the best-quality drinking water available contributes to the optimal production of livestock . Restricted quantity of drinking water or drinking water containing excessive levels of nitrate, TDS, sulfate, and other constituents can affect growth and production of all classes of animals . Drinking-water quality and availability should be evaluated as a cause of poor performance or nonspecific disease conditions in livestock . It is important that attempts to evaluate water quality include obtaining a thorough history, making astute observations, and asking intelligent questions . A thorough laboratory examination of animal specimens and water samples should be evaluated in view of existing standards for livestock drinking-water quality. Curr Biol, 2000 Nov 2, 10(21), R804 - 8 ATP synthase: what dictates the size of a ring? Ferguson SJ. Recent data suggest the source of F(0)F(1) ATP synthase determines a significant and surprising difference in the size of a putative rotating ring of integral membrane subunits of F(0); this can be correlated with biochemical data suggesting there is variation in the number of protons translocated per ATP synthesised. Infect Immun, 2000 Dec, 68(12), 7152 - 5 Neutralizing antibodies to adenylate cyclase toxin promote phagocytosis of Bordetella pertussis by human neutrophils; Weingart CL et al.; A previous study showed that opsonization with human immune serum could either promote or antagonize phagocytosis of Bordetella pertussis by human neutrophils depending on whether the bacteria expressed adenylate cyclase toxin . Opsonization of the wild-type strain inhibited phagocytosis relative to unopsonized controls . In contrast, mutants lacking adenylate cyclase toxin were efficiently phagocytosed when opsonized with human immune serum . In this study, we examined opsonization in the presence or absence of monoclonal antibodies to adenylate cyclase toxin . Addition of neutralizing monoclonal antibodies to adenylate cyclase toxin converted a serum that previously inhibited both attachment and phagocytosis of the wild-type strain to one that increased both attachment and phagocytosis compared to the no-serum control . Monoclonal antibodies that recognize the adenylate cyclase toxin but fail to neutralize activity were without effect . These results suggest that adenylate cyclase toxin inhibits both Fc receptor-mediated attachment and phagocytosis of B . pertussis by neutrophils. Infect Immun, 2000 Dec, 68(12), 6997 - 7002 Identification of mycobacterial surface proteins released into subcellular compartments of infected macrophages; Beatty WL et al.; Considerable effort has focused on the identification of proteins secreted from Mycobacterium spp . that contribute to the development of protective immunity . Little is known, however, about the release of mycobacterial proteins from the bacterial phagosome and the potential role of these molecules in chronically infected macrophages . In the present study, the release of mycobacterial surface proteins from the bacterial phagosome into subcellular compartments of infected macrophages was analyzed . Mycobacterium bovis BCG was surface labeled with fluorescein-tagged succinimidyl ester, an amine-reactive probe . The fluorescein tag was then used as a marker for the release of bacterial proteins in infected macrophages . Fractionation studies revealed bacterial proteins within subcellular compartments distinct from mycobacteria and mycobacterial phagosomes . To identify these proteins, subcellular fractions free of bacteria were probed with mycobacterium-specific antibodies . The fibronectin attachment protein and proteins of the antigen 85-kDa complex were identified among the mycobacterial proteins released from the bacterial phagosome. Infect Immun, 2000 Dec, 68(12), 6826 - 32 Naive human T cells develop into Th1 effectors after stimulation with Mycobacterium tuberculosis-infected macrophages or recombinant Ag85 proteins; Russo DM et al.; Most studies of human T-cell responses in tuberculosis have focused on persons with either active disease or latent infection . Although this work has been critical in defining T-cell correlates of successful versus failed host containment, little is known about the development of Mycobacterium-specific T-cell responses in uninfected persons . To explore this issue, naive T cells from uninfected donors were sensitized in vitro with avirulent Mycobacterium tuberculosis-infected autologous macrophages . T-cell lines primed in this manner proliferated and produced cytokines after challenge with mycobacterial antigens . Of 11 such lines, 8 were high Th1 responders, 2 were low Th1 responders, and 1 was a Th2 responder . Furthermore, similar patterns and magnitudes