Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae Isolated from Vegetables Imported from the Dominican Republic,India, Thailand,and Vietnam

To examine to what extent fresh vegetables imported into Switzerland represent carriers of extended-spectrum-β-lactamase (ESBL)-producing Enterobacteriaceae, 169 samples of different types of fresh vegetables imported into Switzerland from the Dominican Republic, India, Thailand, and Vietnam were analyzed. Overall, 25.4% of the vegetable samples yielded one or more ESBL-producing Enterobacteriaceae, 78.3% of which were multidrug resistant. Sixty isolates were obtained: Escherichia coli, 26; Klebsiella pneumoniae, 26; Enterobacter cloacae, 6; Enterobacter aerogenes, 1; and Cronobacter sakazakii, 1. We found 29 isolates producing CTX-M-15, 8 producing CTX-M-14, 7 producing CTX-M-55, 3 producing CTX-M-65, 1 each producing CTX-M-1, CTX-M-3, CTX-M-27, and CTX-M-63, 5 producing SHV-2, 3 producing SHV-12, and 1 producing SHV-2a. Four of the E. coli isolates belonged to epidemiologically important clones: CTX-M-15-producing B2:ST131 (1 isolate), D:ST405 (1 isolate), and D:ST38 (2 isolates). One of the D:ST38 isolates belonged to the extraintestinal enteroaggregative E. coli (EAEC) D:ST38 lineage. Two of the K. pneumoniae isolates belonged to the epidemic clones sequence type 15 (ST15) and ST147. The occurrence of antibiotic-resistant pathogenic and commensal Enterobacteriaceae in imported agricultural foodstuffs constitutes a source of ESBL genes and a concern for food safety.     

Cloning, sequencing, expression, and antigenic characterization of rMSP4 from Anaplasma marginale isolated from Paraná State, Brazil

Anaplasmosis is a bovine intraerythrocytic disease caused by the bacterium Anaplasma marginale; it causes significant economic losses in tropical and subtropical regions, worldwide. The msp4 gene of an A. marginale strain isolated in Paran , Brazil, was amplified by PCR and sequenced; its cloning into the pET102/D-TOPO vector produced an msp4-6xHis-V5-HP thioredoxin fusion gene construct. This recombinant clone was over-expressed in Escherichia coli BL21(DE-3); the expressed fusion protein was found almost entirely in the insoluble form (inclusion bodies) in the cell lysate. The inclusion bodies were solubilized with urea and the recombinant protein was purified by Ni-NTA column and dialyzed. This method produced a relatively high yield of rMSP4, which was used to immunize rabbits. The deduced amino acid sequence encoded by MSP4 showed 99% homology to A. marginale isolates from Florida, USA, and from Minas Gerais, Brazil. Both rMSP4 and native MSP4 were recognized by post-immunization rabbit serum, showing that rMSP4 has conserved epitopes. As antigenicity was preserved, rMSP4 might be useful for the development of vaccine against anaplasmosis.     

If good things come from above,do bad things come from below?

Factors in the synovial fluid that maintain healthy articular cartilage, such as hyaluronic acid and lubricin, come from above. Is it possible that factors which lead to the destruction of cartilage come from below in the subchondral bone? The recent acquisition of tools to probe early events in osteoarthritis is shedding new light on possible contributions from this compartment on the initiation and progression of the disease. Tanamas and co-workers now provide evidence that bone marrow lesions in the subchondral bone are predictive, both of loss of cartilage and of formation of subchondral cysts. These data provoke questions about the nature and role of bone marrow lesions.Finding the factors that initiate, or the mechanisms that lead to progression of, osteoarthritis (OA) has proven frustrating and largely unproductive. Identification of risk factors for the condition - such as prior trauma to the joint, elevated body weight and female sex - may have helped with management of OA but has done little to progress understanding of the underlying factors that drive it. OA research has been more difficult than research for some other diseases of the skeleton, for several important reasons. Early OA, at the level of symptoms, can be episodic, making it difficult to identify the disease and to follow it longitudinally. Since the main early symptom is pain, clinical trials of new therapies have been problematic. Animal experiments have been bedevilled by a lack of models that accurately replicate the human disease. And perhaps, as argued by a minority of workers in the field, disease initiators have been sought in the wrong place; that is, cartilage versus bone.The recent study of Tanamas and colleagues highlights the way in which new-generation imaging holds the promise of shedding new light on this old problem [1]. In particular, high-resolution magnetic resonance imaging (MRI) can now deliver objective, measurable information about all structures of the joint, including the amount and quality of articular cartilage, and is also a powerful tool to investigate the subchondral bone. The holy grail of clinical investigation, namely longitudinal study with quantitative endpoints, is now accessible for OA. What Tanamas and colleagues'' study shows is important because it adds to emerging evidence that processes in the subchondral bone relate strongly to changes in the volumetric amount of articular cartilage. Specifically, bone marrow lesions (BMLs), the mysterious MRI-bright regions in the subchondral bone that occur more commonly in OA, were shown to be predictive of loss of cartilage and of formation of subchondral cysts. In turn, cysts were more likely than BMLs to occur in association with loss of cartilage.These data pose the intriguing question of whether BMLs encode key clues to the aetiology of OA. Longitudinal studies have shown that the presence of BMLs constitutes a potent risk factor for structural deterioration in knee OA [2]. BML enlargement has been strongly associated with increased cartilage loss, and Tanamas and colleagues'' data further suggest that their conversion into cysts is even more predictive of cartilage loss. Significantly, a reduction in the extent of BMLs on MRI has been shown to associate with a decrease in cartilage degradation [3]. Since the origin of BMLs is not known, its investigation needs to be prioritised as an important research topic. Current informed guesses are that BMLs comprise regions of oedema, perhaps secondary to episodes of local ischaemia. Although it is not possible to biopsy BMLs in patients with early OA, several studies have sought to correlate the MRI findings with histology in more severe disease. Regions of BMLs in end-stage OA patients at knee replacement were more likely to exhibit oedema, bone necrosis and trabecular abnormalities than were control sites [4].If BMLs are secondary to local ischaemia in the subchondral bone, there are several possible consequences. Firstly, the supply of nutrients and oxygen from regions of ischaemic subchondral bone, to the overlying articular cartilage, might be reduced. Cartilage nutrition has been considered to derive from the synovial fluid. The work of Imhof and colleagues, however, suggested that more than 50% of the glucose, oxygen and water requirements of cartilage are provided by perfusion from the subchondral vessels [5]. They described the dense subchondral vasculature in close proximity to the cartilage, and the micro-channels that penetrate the subchondral mineralisation zone and permit communication between the bone and the cartilage. More recent work indicates that small molecules can diffuse, in healthy joints, bidirectionally from the synovial compartment into the cartilage and underlying bone and from the subchondral bone into the overlying cartilage [6]. Inspection of the osteochondral junction of long bones reveals that osteocytes and osteocyte canaliculi, which are also probable conduits of nutrients, are intimately associated with the articular cartilage. Experimental interruption of contact between articular cartilage and subchondral bone results in degeneration of the cartilage, and osteoblasts from OA subchondral bone conferred catabolic changes in articular chondrocytes [7].Secondly, osteocyte death in bone is becoming recognised as a signalling event for osteoclastic removal of the nonviable bone and its replacement in a remodelling episode [8]. Although subchondral bone is constantly being remodelled, concentration of this activity in a particular region of the bone could alter its mechanical integrity and its ability to properly support the overlying cartilage.Tanamas and colleagues conclude that cysts (and BMLs) may provide therapeutic targets for the treatment of knee OA [1]. Certainly, the recent acquisition of tools to probe early events in subchondral bone in OA should deliver rapid advances in our understanding of the natural history of this condition.     

Immunomodulatory compounds from Pestalotiopsis leucothës, an endophytic fungus from Tripterygium wilfordii

The immunomodulatory effects of three compounds designated BS, GS, and YS produced by Pestalotiopsis leucothës, an endophytic fungus isolated from Tripterygium wilfordii, were evaluated. The 50% inhibition concentration (IC₅₀) value of BS in the proliferative assay with various stimulating agents such as phytohemagglutinin-M (PHA-M), phorbol myristate acetate (PMA)/ionomycin, mixed lymphocyte reaction (MLR) and poke weed mitogen (PWM) was 0.35, 1.6, 0.8 and 5.4 μg/ml, respectively. In addition, BS significantly inhibited the production of cytokines such as interleukin (IL)-1β, IL-2, interferon (IFN)-γ and tumor necrosis factor (TNF)-α, by peripheral blood mononuclear cells (PBMNC) and soluble IL-2 receptor expression at concentrations greater than 1 μg/ml. Inhibition of PHA stimulated PBMNC proliferation and IL-2 and sIL-2R production by BS indicates that it is a T-cell specific immunosuppressant. However, BS also moderately inhibited immunoglobulin (Ig) G and M at concentrations greater than 1 μg/ml suggesting that it also has B cell immunosuppressive effects. YS was 10% less active than BS in all assay systems. In contrast, GS exhibited both suppression and enhancement of PBMNC proliferation in the presence of various stimulants. However, GS inhibited PWM stimulated PBMNC proliferation and IL-4 and IgG and IgM production at concentrations above 1 μg/ml. All three fungal compounds altered the percentage of T-lymphocyte subpopulations only at high concentrations. Cell viability was not affected at the immunosuppressive concentrations of these compounds. In conclusion, work from our laboratory has identified three potentially potent immunomodulatory compounds from P. leucothës. These compounds have variable effects on T- and B-cells and monocytes. They may partially explain the immunosuppressive activity of T. wilfordii. In addition, they may represent a new source of immunomodulatory compounds for the treatment of human immune mediated diseases.     

Pheno- and genotyping of Staphylococcus aureus, isolated from bovine milk samples from São Paulo State, Brazil

In the present study, 87 Staphylococcus aureus isolates obtained from milk samples of 87 cows with mastitis in 6 different municipal districts of 2 regions of Sao Paulo State, Brazil, were compared pheno- and genotypically. Pulsed-field gel electrophoresis (PFGE) analysis of the strains was performed, and PCR was carried out to detect genes for a number of staphylococcal cell surface proteins, exoproteins, and 3 classes of agr genes. Nine distinct S. aureus lineages (LA-LI) were identified by PFGE. The lineages LA and LE, which accounted together for 63 strains (72.2%), were prevalent and had been collected from all of the 6 municipal districts, indicating a broad geographic distribution of these lineages; LB, LC, LD, LF, LG, LH, and LI, however, were isolated sporadically and accounted for 24 strains (27.8%). Some characteristics, like penicillin resistance and the presence of cap8 and agr class II genes, were associated with the prevalent lineages (LA and LE), and penicillin susceptibility and the presence of cna and cap5 genes were associated with sporadic lineages. According to the present results, some S. aureus lineages possess a combination of genes that confer the propensity to cause and disseminate infection, and only a limited number of clones are responsible for the cases of bovine mastitis on the various farms.     

Production,purification, and properties of β-glucosidase from Candida wickerhamii

Summary Candida wickerhamii growing on cellobiose produced -glucosidase with high activity against -nitrophenyl glucoside (PNPG) but low activity against cellobiose. -glucosidase production was constitutive, and was repressed by -glucosides and glucose. -glucosides containing an aromatic moiety in the aglycon were the best substrates for -glucosidase indicating that the enzyme is an aryl--glucosidase. A -glucosidase from C. wickerhamii cells was purified by (NH₄)₂SO₄ precipitation, dialysis, ion-exchange chromatography and gel filtration. The purified enzyme was homogeneous as shown by sodium-dodecyl-sulphate polyacrylamide gel electrophoresis and discontinuous gel electrophoresis. The purified enzyme hydrolysed PNPG but not cellobiose. The K_m of the enzyme was 0.185 mM. Glucose inhibited the enzyme competitively and the K_i was 7.5 mM. The apparent molecular mass was 97,000. The optimum pH and temperature for enzyme activity were between pH 7 and 7.4 and 40°C respectively. At temperatures of 45°C and greater the enzyme was inactivated. The activation energy of the enzyme was 29.4 kJ · mol^-1.     

Separation,Characterization, and Specificity of α-Mannosidases from Vigna umbellata

Information about the specificity of glycosidase enzymes is important since it affects their use for characterization and synthesis of oligosaccharides. Two α-mannosidases (EC 3.2.1.24), I and II, were isolated from rice beans (Vigna umbellata). The native molecular weight of both isozymes was estimated to be 329,000, but pIs of form I were 5.03-5.34 and pIs of form II were 5.46-6.20. The two isozymes were characterized in terms of optimal pH and temperature, effects of metal ions, inhibition by swainsonine and 1-deoxymannojirimycin, and kinetic parameters for p-nitrophenyl-α-D-mannopyranoside and Manα(1-2)Man. Both enzymes were more specific towards Manα(1-2)Man in both hydrolysis and synthesis, but their hydrolytic specificities towards Manα(1-3)[Manα(1-6)]Man were different.     

HIV and HCV： from Co-infection to Epidemiology, Transmission, Pathogenesis, and Treatment

Human immunodeficiency virus (HIV) is the infectious agent causing acquired immu-nodeficiency syndrome (AIDS),a deadliest scourge of human society. Hepatitis C virus (HCV) is a major causative agent of chronic liver disease and infects an estimated 170 million people worldwide,resulting in a serious public health burden. Due to shared routes of transmission,co-infection with HIV and HCV has become common among individuals who had high risks of blood exposures. Among hemophiliacs the co-infection rate accounts for 85%; while among injection drug users (IDU) the rate can be as high as 90%. HIV can accelerate the progression of HCV-related liver disease,particularly when immunodeficiency has developed. Although the effect of HCV on HIV infection is controversial,most studies showed an increase in mortality due to liver disease. HCV may act as a direct cofactor to fasten the progression of AIDS and decrease the tolerance of highly active antiretroviral therapy (HARRT). Conversely,HAART-related hepatotoxicity may enhance the progression of liver fibrosis. Due to above complications,co-infection with HCV and HIV-1 has imposed a critical challenge in the management of these patients. In this review,we focus on the epidemiology and transmission of HIV and HCV,the impact of the two viruses on each other,and their treatment.     

Abstracts from “Neirokhimiya,” Vol. 17, No. 1, 2000

Abstract List

Abstracts from Neirokhimiya, Vol. 17, No. 1, 2000     

Phylloquinone, what can we learn from plants?

The plant plasma membrane contains redox proteins able to mediate a trans-membrane electron flow. This electron flow might be responsible for the generation of the active oxygen species observed as a reaction to pathogen attack or stress. Vitamin K1 could be identified as a possible lipid soluble electron carrier in plant plasma membrane preparations. Such a function would be analogous to coenzyme Q in animal plasma membranes. What we are going to outline in this contribution is a concept of how the electron transport system of the plant plasma membrane could interact with quinones, thus contributing to the metabolism of free radicals in plants.     

Hermosillol,an 8,3′-neolignan from Krameria sonorae

The aerial parts of Krameria sonorae afforded seven known neolignans and nor-neolignans, and as a major component, a new 8,3′-neolignan which was named hermosillol.     

Harmalidine,A β-carboline alkaloid from peganum harmala

A new β-carboline alkaloid, harmalidine, has been isolated from the seeds of Peganum harmala and its structure elucidated by spectral and chemical studies.     

Seroprevalence and isolation of Toxoplasma gondii from sheep from São Paulo state, Brazil

Sheep are important in the epidemiology of Toxoplasma gondii infection but little is known of ovine toxoplasmosis in Brazil. Antibodies to T. gondii were assayed in sera of 495 sheep from 36 counties of S?o Paulo State, Brazil, using the modified agglutination test (MAT titer > or =1:25) and found in 120 (24.2%). Samples of brain, heart, and diaphragm of 82 seropositive sheep were pooled, digested in pepsin, and bioassayed in mice. Toxoplasma gondii was isolated from tissue homogenates of 16 sheep and the isolates were designated TgShBr1-16. Six of the 16 T. gondii isolates killed 100% of infected mice. Results indicate that asymptomatic sheep can harbor mouse-virulent T. gondii, and hence they can serve as a source of infection for humans.     

A new glossiphoniid leech from Catemaco Lake, Veracruz, México

Haementeria acuecueyetzin n. sp. from Catemaco Lake, Veracruz, Mexico, is described based on the examination of 6 specimens. This new hematophagous leech species resembles other members of the genus in the number and position of the eyespots, number of compact salivary glands, and in the presence of 2 pairs of spheroidal mycetomes, but it is distinguished from the other species by having 6 rows of longitudinal smooth white papillae in the dorsal surface and numerous tubercles in dorsal and ventral surfaces. This new species represents the third species of Haementeria in the Northern Hemisphere of the Americas.     

Are Large, Infrequent Disturbances Qualitatively Different from Small, Frequent Disturbances?

In this article, we develop a heuristic model of ecosystem-disturbance dynamics that illustrates a range of responses of disturbance impact to gradients of increasing disturbance extent, intensity, or duration. Three general kinds of response are identified and illustrated: (a) threshold response, (b) scale-independent response, and (c) continuous response. Threshold responses are those in which the response curve shows a discontinuity or a sudden change in slope along the axis of increasing disturbance extent, intensity, or duration. The response threshold occurs at a point where the force of the disturbance exceeds the capacity of internal mechanisms to resist disturbance, or where new mechanisms of recovery become involved. Within this conceptual framework, we find that some unusually large or intense disturbances, but not all, produce qualitatively different responses compared with similar disturbances of lesser magnitude. If disturbance impact does not increase with increasing disturbance extent, intensity, or duration, or if the response curve changes monotonically, then large disturbances are not qualitatively different from small ones. For example, jack pine tends to become reestablished after stand-replacing fire in boreal forests, regardless of fire size, because its serotinous cones provide an adequate seed source throughout the burned area. Thus, large fires are not qualitatively different from small fires in terms of jack pine reproduction. However, if disturbance impact does increase abruptly at some point with increasing disturbance extent, intensity, or duration, often because of thresholds in the capacity of internal mechanisms to resist or respond to disturbance impact, then large disturbances are qualitatively different from small ones, at least for some parameters of ecological response. For example, balsam fir and white cedar can recolonize a small burned patch of boreal forest in close proximity to surviving individuals of these species, but they will be eliminated from a large burn because of their susceptibility to fire-caused mortality and their inability to disperse their seeds over long distances. The conceptual framework presented here permits some new insights into the dynamics of natural systems and may provide a useful tool with which managers can assess the potential for catastrophic damages resulting from large, infrequent disturbances. Received 14 July 1998; accepted 29 September 1998     

Grass,mosses, algae,or leaves? Food preference among shredders from open-canopy streams

Shredder feeding is a vital process in making decomposition products available to biota in streams. To investigate which food sources shredders in open-canopy streams exploit, we conducted a feeding preference experiment with the invertebrate detritivores Limnephilus bipunctatus and Nemoura sp., which are commonly found in open-canopy streams on the Swedish island of Öland in the southern Baltic Sea. Leaves of birch, Swedish whitebeam, and shrubby cinquefoil; dead and fresh grass; water moss; and algae were offered to the shredders in multi- and single treatments. We hypothesized that food with high nutritional value would be preferred. Both taxa preferred leaves of shrubby cinquefoil, a bush common in the riparian zone of Öland streams; additionally Nemoura sp. also chose algae. Dead grass, the most abundant food source in the streams during the whole year, was the least consumed food type. The fresh food types had highest nutritional value, measured as carbon to nitrogen content. Therefore, food quality could not alone explain the preference of shrubby cinquefoil. However, among the detritus type offered, shrubby cinquefoil had the highest nutritional value. Shrubby cinquefoil may constitute one important energy source to these open-canopy stream ecosystems and may be essential in maintaining an abundant shredder community in these streams. Thus, the results of this study indicate that detrital resources are indeed important in open-canopy stream systems.     

Ichthyofauna from wetlands of San Pedro, Balancán, Tabasco, México

San Pedro River's wetlands sustain trophic nets in the fluvial system, due to the high habitat availability, and space and temporal variations. In order to describe the relationship between environmental parameters and ichthyofauna, this study evaluated fish assemblages composition, distribution, abundance, density, biomass, richness species, diversity and equitability in the wetlands. Sampling considered three different sites and climatic seasons (dry, rainy and cold fronts). The physical and chemical parameters considered were dissolved oxygen, temperature, pH, water transparency and the depth. Fishes were caught with a shrimp net, after six minutes towings and were identified afterwards. A total of 1 049 organisms of 25 fish species were caught, two of which were exotic species: Oreochromis niloticus and Parachromis managuense. A total of 23 species were found at site I (with the highest density 0.23 ind./m2), 17 at site II (0.23 ind./m2) and 14 at site III (0.12 ind./m2). The dry season had the highest species number with 21 species, followed by the rainy season with 17 species, and the cold season with five species. Similarly, the highest biomass (8.30 g/m2) was found in dry season, followed by the rainy (2.16 g/m2) and the cold seasons (0.03 g/m2). Considering seasons, highest density was found during the dry (0.436 ind./m2), followed by the rainy (0.213 ind./m2) and the cold (0.023 ind./m2) seasons. The dominant density species during the study period, according to the quadrants graphic of Olmstead-Tukey were: Petenia splendida, Vieja heterospila, Vieja synspila, Dorosoma petenense and Astyanax aeneus. There were significant differences in the species richness among sites. Temperature, depth and transparency showed differences among the seasons. The canonical correspondence analysis indicated that fish distribution was governed by environmental parameters during all seasons. In terms of fish abundance and composition, environmental parameters play an important role showing spatial and temporal differences in the ecosystem, this could be explained with the fact that most of young fishes have a movement behavior to the wetlands, searching refuge and feed during the dry season. Considering the diversity indexes variation, it may be concluded that San Pedro River's wetlands correspond to a system where the ichthyofauna composition fluctuates spatial and seasonally.     

Campanian–early Eocene cephalopods from Kharga Oasis,Western Desert,Egypt

The Campanian–lower Eocene sedimentary succession in the Kharga Oasis yields rare cephalopods that have so far received little attention. Eight cephalopod species; six nautiloids and two ammonites, are identified in the study area. The nautiloids are referred to five genera in three families. All nautiloid species are recorded from the Paleocene and Eocene rocks, two of which are described as new, as follows: Cimomia kurkurensis nov. sp. and Deltoidonautilus hassani nov. sp. The two ammonite species are Libycoceras ismaelis (von Zittel, 1884) and Baculites ovatus Say, 1820, representing the families Sphenodiscidae and Baculitidae, respectively. Baculites anceps of Quaas (1902, non Lamarck, 1822) from the Maastrichtian of the Western Desert of Egypt is here assigned to Baculites ovatus. The palaeobiogeography of these species is studied in detail.