High Prevalence of Vancomycin-Resistant Enterococci in Swedish Sewage

In Europe the use of the growth promoter avoparcin is considered to have selected for vancomycin-resistant enterococci (VRE). Sweden ceased using avoparcin in 1986, and only occasional cases of VRE from hospitals have been reported since 1995. Within the framework of a European study, samples from urban raw sewage, treated sewage, surface water, and hospital sewage in Sweden (n = 118) were screened for VRE. Surprisingly, VRE were isolated from 21 of 35 untreated sewage samples (60%), from 5 of 14 hospital sewage samples (36%), from 6 of 32 treated sewage samples (19%), and from 1 of 37 surface water samples. Thirty-five isolates from 33 samples were further characterized by geno- and phenotyping, MIC determination, and PCR analysis. Most isolates (30 of 35) carried the vanA gene, and the majority (24 of 35) of the isolates were Enterococcus faecium. Most of the VRE were multiresistant. The typing revealed high diversity of the isolates. However, one major cluster with seven identical or similar isolates was found. These isolates came from three different sewage treatment plants and were collected at different occasions during 1 year. All VRE from hospital sewage originated from one of the two hospitals studied. That hospital also had vancomycin consumption that was 10-fold that of the other. We conclude that VRE were commonly found in sewage samples in Sweden. The origin might be both healthy individuals and individuals in hospitals. Possibly, antimicrobial drugs or chemicals released into the sewage system may sustain VRE in the system.     

Temperature-dependent structural transition following X-ray-induced metal center reduction in oxidized cytochrome c oxidase

Cytochrome c oxidase (CcO) is the terminal enzyme in the electron transfer chain in the inner membrane of mitochondria. It contains four metal redox centers, two of which, Cu_B and heme a₃, form the binuclear center (BNC), where dioxygen is reduced to water. Crystal structures of CcO in various forms have been reported, from which ligand-binding states of the BNC and conformations of the protein matrix surrounding it have been deduced to elucidate the mechanism by which the oxygen reduction chemistry is coupled to proton translocation. However, metal centers in proteins can be susceptible to X-ray-induced radiation damage, raising questions about the reliability of conclusions drawn from these studies. Here, we used microspectroscopy-coupled X-ray crystallography to interrogate how the structural integrity of bovine CcO in the fully oxidized state (O) is modulated by synchrotron radiation. Spectroscopic data showed that, upon X-ray exposure, O was converted to a hybrid O∗ state where all the four metal centers were reduced, but the protein matrix was trapped in the genuine O conformation and the ligands in the BNC remained intact. Annealing the O∗ crystal above the glass transition temperature induced relaxation of the O∗ structure to a new R∗ structure, wherein the protein matrix converted to the fully reduced R conformation with the exception of helix X, which partly remained in the O conformation because of incomplete dissociation of the ligands from the BNC. We conclude from these data that reevaluation of reported CcO structures obtained with synchrotron light sources is merited.     

Cell proliferation and gill morphology in anoxic crucian carp

Is DNA replication/cell proliferation in vertebrates possible during anoxia? The oxygen dependence of ribonucleotide reductase (RNR) could lead to a stop in DNA synthesis, thereby making anoxic DNA replication impossible. We have studied this question in an anoxia-tolerant vertebrate, the crucian carp (Carassius carassius), by examining 5'-bromo-2'-deoxyuridine incorporation and proliferating cell nuclear antigen levels in the gills, intestinal crypts, and liver. We exposed crucian carp to 1 and 7 days of anoxia followed by 7 days of reoxygenation. There was a reduced incidence of S-phase cells (from 12.2 to 5.0%) in gills during anoxia, which coincided with a concomitant increase of G(0) cells. Anoxia also decreased the number of S-phase cells in intestine (from 8.1 to 1.8%). No change in the fraction of S-phase cells ( approximately 1%) in liver was found. Thus new S-phase cells after 7 days of anoxia were present in all tissues, revealing a considerable rate of DNA synthesis. Subsequently, the oxygen-dependent subunit of crucian carp RNR (RNRR2) was cloned. We found no differences in amino acids involved in radical generation and availability of the iron center compared with mouse, which could have explained reduced oxygen dependence. Furthermore, the amount of RNRR2 mRNA in gills did not decrease throughout anoxia exposure. These results indicate that crucian carp is able to sustain some cell proliferation in anoxia, possibly because RNRR2 retains its tyrosyl radical in anoxia, and that the replication machinery is still maintained. Although hypoxia triggers a 7.5-fold increase of respiratory surface area in crucian carp, this response was not triggered in anoxia.     

中药王不留行化学成分的研究

从河北产麦蓝菜(Vaccaria segetalis(Neck)Garcke.)的种子王不留行中分离得到5个化合物,经波谱分析和化学方法分别鉴定为:Tetracosanoic acid(1),E-3-phenyl-2-Propenoic acid(2),stigmast-7,22-dien-3-ol(3),stigmast-7,22-dien-3-ol 3β-O-[β-D-glucopy-ranoside](4),9,12-octadecadienoic acid N-(2-hydroxyethyl)(5)。以上各化合物都是首次从该植物中分得。化合物5为首次从天然界分得。     

Mechanism and Factors Influencing the Veronal Inhibition of Bacterial Spore Germination

The inhibitory effect of sodium 5,5-diethyl barbiturate (Veronal) on the L-alanine-induced initiation of germination of Bacillus subtilis spores was examined. Veronal reversibly inhibited the initiation of germination by a noncompetitive mechanism. The inhibition was time-independent and it took place whether L-alanine was or was not allowed to permeate the spore before the addition of the inhibitor. The concentration of the inhibitor and the pH of the initiation system were important factors determining the effectiveness of Veronal as an inhibitor. The magnitude of the inhibition increased linearly with decreasing pH at constant concentration and with increasing concentration at constant pH. These results suggest that the inhibition involves a permeability phenomenon related to the access of drug to the active sites in the spore and that the entry of Veronal into the spores is regulated by the concentration of undissociated molecule. At the physiologically important pH of 7.4, initiation with alanine in phosphate buffer at high spore densities (about 10(9) spores per ml) was 50% inhibited by 4 mM Veronal, and 8mM Veronal inhibited initiation completely. L-Alanine initiation in tris(hydroxymethyl)amino-methane-hydrochloride buffer was completely inhibited by 5 mM Veronal. The inhibition could be partially reversed by the combined addition of D-fructose, D-glucose, and K(+). Possible reasons for the failure of otherwise inhibitory concentrations of Veronal to inhibit completely the L-alanine-induced initiation when a combination of fructose, glucose, and K(+) was present and a suggested relationship to two functional roles of L-alanine in the initiation of germination are discussed.     

Root distribution and water uptake patterns of maize cultivars field-grown under differential irrigation

Summary Rooting and water uptake patterns were determined for three maize (Zea mays L) varieties field-grown during the 1983/84 dry season under seven irrigation levels on a sandy loam soil. Roots were mainly concentrated in the top 22 cm due to a 40 cm thick compact gravelly layer occurring from about this depth in the profile. There were significant varietal differences, distinguished by root length density (RLD) and length/weight ratio (LAR) distributions at depth and at varying soil moisture regimes. These properties were related to water extraction patterns and grain yields. Yields obtained at adequate soil moisture were 6.9 tha⁻¹ for TZESR-W (var 1), 4.2 t/ha for TZSR-W (var 2) and 3.7t ha⁻¹ for FARZ-7 (var 3). These yeilds were respectively associated with maximum RLD of 2.56, 1.88 and 1.70 cm cm⁻³ and corresponding LWR of 2.64, 1.93 and 1.62 cm mg⁻¹. Average seasonal water uptake was estimated at 4.2, 3.0 and 2.8 mm day⁻¹ for var 1, 2 and 3, respectively. Better performance of var 1 was attributed to the development of a more active and deep rooting system.     

Site-specific phosphorylation of Tau protein is associated with deacetylation of microtubules in mouse spermatogenic cells during meiosis

Tau is one of the microtubule-associated proteins and a major component of paired helical filaments, a hallmark of Alzheimer’s disease. Its expression has also been indicated in the testis. However, its function and modification in the testis have not been established. Here, we analyzed the dynamics of phosphorylation patterns during spermatogenesis. The expression of Tau protein and its phosphorylation were shown in the mouse testis. Immunohistochemistry revealed that the phosphorylation was strongly detected during meiosis. Correspondingly, the expression of acetylated tubulin was inversely weakened during meiosis. These results suggest that phosphorylation of Tau protein contributes to spermatogenesis, especially in meiosis.     

High Prevalence,Coinfection Rate,and Genetic Diversity of Retroviruses in Wild Red Colobus Monkeys (Piliocolobus badius badius) in Ta? National Park,C?te d'Ivoire

Simian retroviruses are precursors of all human retroviral pathogens. However, little is known about the prevalence and coinfection rates or the genetic diversity of major retroviruses—simian immunodeficiency virus (SIV), simian T-cell lymphotropic virus type 1 (STLV-1), and simian foamy virus (SFV)—in wild populations of nonhuman primates. Such information would contribute to the understanding of the natural history of retroviruses in various host species. Here, we estimate these parameters for wild West African red colobus monkeys (Piliocolobus badius badius) in the Taï National Park, Côte d''Ivoire. We collected samples from a total of 54 red colobus monkeys; samples consisted of blood and/or internal organs from 22 monkeys and additionally muscle and other tissue samples from another 32 monkeys. PCR analyses revealed a high prevalence of SIV, STLV-1, and SFV in this population, with rates of 82%, 50%, and 86%, respectively. Forty-five percent of the monkeys were coinfected with all three viruses while another 32% were coinfected with SIV in combination with either STLV or SFV. As expected, phylogenetic analyses showed a host-specific pattern for SIV and SFV strains. In contrast, STLV-1 strains appeared to be distributed in genetically distinct and distant clades, which are unique to the Taï forest and include strains previously described from wild chimpanzees in the same area. The high prevalence of all three retroviral infections in P. b. badius represents a source of infection to chimpanzees and possibly to humans, who hunt them.Lentiviruses and deltaretroviruses that infect African nonhuman primates have received considerable attention as they are the precursors of all pathogenic human retroviruses: human immunodeficiency virus types 1 and 2 (HIV-1/HIV-2) and human T-cell lymphotropic virus type 1 (HTLV-1). These human infections are the results of past zoonotic transfers of simian immunodeficiency virus (SIV) and simian T-cell lymphotropic viruses type 1 (STLV-1) from wild monkeys and apes into local human populations, presumably through primate hunting and handling of primate bushmeat (13, 19, 43, 46, 55, 58, 59). Via the same route, zoonotic transmission of simian foamy virus (SFV), a spumaretrovirus whose exact pathogenicity in human hosts is still unknown, has also been shown (64). The increasing contact between humans and wild primates implies that further zoonotic transmission of retroviruses is likely to happen (42, 63). Studying the occurrence and circulation of simian retroviruses such as SIV, STLV-1, and SFV in wild primate populations enables us to better understand retrovirus evolution in primates and also provides tools for monitoring possible future retroviral zoonotic events.Systematic studies of SIV, STLV-1, and SFV in wild primates are relatively rare. Many use bushmeat samples, which can vary in their quality and are prone to cross-contamination from butchering and storage with other carcasses. Confiscated primates are also not representative of the situation in the wild since the animals are caught at a young age when the occurrence of different retroviruses may be extremely low (24). The technical possibilities for the detection of various pathogens in noninvasive samples such as urine and feces have greatly improved and are frequently used; however, in general, the sensitivity of detection methods is higher when blood and tissue samples are used (25, 32, 47). Such samples can be collected if fresh carcasses are found, or they can be collected by anesthetizing live primates for sampling purpose, animal translocation, or medical intervention, such as snare removal. The practical and ethical issues of each of the sampling methods have been discussed elsewhere (12, 14).Red colobus monkeys [Procolobus (Piliocolobus)] are interesting subjects for retroviral infection studies for a number of reasons. First, they are widely distributed (yet in a fragmented manner) from East to West Africa, which suggests that red colobus species and subspecies, or more likely ancestor(s) of these, could have been key hosts in transmitting retroviruses across tropical Africa (4, 54). Second, as they are herbivore primates, the hunting of other primates can be excluded as a route of infection. Finally, these monkeys are frequently hunted by humans and chimpanzees and represent a possibly large reservoir for retroviruses and other pathogens that ought to be investigated further (2, 45).Very little information is available about the prevalence and coinfection of SIV, STLV-1, and SFV in wild red colobus monkeys across Africa. In other colobine monkeys only SIV has been documented: in olive colobus (Procolobus verus) in Côte d''Ivoire and in black and white colobus (Colobus guereza) in Cameroon (7, 8). Based on fecal samples from habituated adult individuals, the prevalence of SIV in West African red colobus monkeys (SIVwrc; local subspecies, Piliocolobus badius badius) has been estimated to a minimum of 26% in the Taï National Park, Côte d''Ivoire, but the authors recognized the low sensitivity of viral RNA detection in fecal samples (34). Another study conducted on the same population revealed that 5 out of 10 blood samples were SIV positive (7). These results highlight that the most reliable prevalence data are based on analyses of blood/tissue samples although such sampling is not always feasible for reasons discussed above. Published prevalence information concerning STLV-1 and SFV in wild red colobus monkeys (STLV-1wrc and SFVwrc) in the same area is restricted to results obtained from analyses of a limited number of blood and necropsy samples collected as a part of studies whose focus was on cross-species transmission of these two viruses to chimpanzees (27, 28). However, these samples indicated a high prevalence of STLV-1wrc and SFVwrc in the red colobus monkey population (56% and 90%, respectively). A recent study from Uganda, East Africa, estimated the prevalence of SIV, STLV-1, and SFV in another red colobus species (Piliocolobus rufomitratus tephrosceles) to be 22.6%, 6.4%, and 97%, respectively (15). The study was performed using blood samples collected from anesthetized wild red colobus monkeys living in their natural habitat, which allowed reliable assessment of the prevalence and genetic diversity of these three retroviruses.The preliminary data from the Taï National Park indicate that there might be great variation in the prevalence of retroviruses across the African continent, even in closely related species of wild primates. Here, we aimed at generating reliable prevalence and coinfection data for SIVwrc, STLV-1wrc, and SFVwrc based on the analysis of blood and tissue samples from wild Western red colobus monkeys. We expected that this would allow for proper comparison of retroviral prevalence in the allied species P. b. badius and P. r. tephrosceles.