Long-term fermented soybean paste improves metabolic parameters associated with non-alcoholic fatty liver disease and insulin resistance in high-fat diet-induced obese mice

Recently, Korean traditional fermented soybean paste, called Doenjang, has attracted attention for its protective effect against diet-related chronic diseases such as obesity and type 2 diabetes. Long-term fermented soybean pastes (LFSPs) are made by fermentation with naturally-occurring microorganisms for several months, whereas short-term fermented soybean pastes (SFSPs) are produced by shorter-time fermentation inoculated with a starter culture. Here, we demonstrate that administration of LFSP, but not SFSP, protects high-fat diet (HFD)-fed obese mice against non-alcohol fatty liver disease (NAFLD) and insulin resistance. LFSP suppressed body weight gain in parallel with reduction in fat accumulation in mesenteric adipose tissue (MAT) and the liver via modulation of MAT lipolysis and hepatic lipid uptake. LFSP-treated mice also had improved glucose tolerance and increased adiponectin levels concomitantly with enhanced AMPK activation in skeletal muscle and suppressed expression of pro-inflammatory cytokines in skeletal muscle and the liver. LFSP also attenuated HFD-induced gut permeability and lowered serum lipopolysaccharide level, providing an evidence for its probiotic effects, which was supported by the observation that treatment of a probiotic mixture of LFSP-originated Bacillus strains protected mice against HFD-induced adiposity and glucose intolerance. Our findings suggest that the intake of LFSP, but not SFSP, offers protection against NAFLD and insulin resistance, which is an effect of long-term fermentation resulting in elevated contents of active ingredients (especially flavonoids) and higher diversity and richness of Bacillus probiotic strains compared to SFSP.     

Plant biodiversity patterns along a climatic gradient and across protected areas in West Africa

Knowledge of spatial patterns of biological diversity is fundamental for ecological and biogeographical analyses and for priority setting in nature conservation, particularly in West Africa where the existing high biodiversity is increasingly threatened by human activities. The maximum entropy approach was used to model the geographic distribution of 3,393 vascular plant species at a spatial resolution of 0.0833°. Species richness decreases along temperature and precipitation gradients with high species numbers in the south and lower numbers towards the north of the transect. All centres of plant species diversity are confined to humid areas in concordance with the high positive correlation between species richness and rainfall which appears to be the most important delimiter for the distribution ranges of many species in the area. The effectiveness of the existing protected areas at regional and national levels is investigated based on the proportion of species covered. Considering the whole study area, 95% of all species are covered by protected areas according to their distribution ranges. However, the proportion of species covered is considerably lower for some countries such as Benin and Togo. Our results could provide guidance for essential land use management interventions to decision‐makers and conservationists in the region.     

Mechanistic studies of sesquiterpene cyclases based on their carbon isotope ratios at natural abundance

During the process of terpene biosynthesis, C–C bond breaking and forming steps are subjected to kinetic carbon isotope effects, leading to distinct carbon isotopic signatures of the products. Accordingly, carbon isotopic signatures could be used to reveal the ‘biosynthetic history’ of the produced terpenoids. Five known sesquiterpene cyclases, regulating three different pathways, representing simple to complex biosynthetic sequences, were heterologously expressed and used for in vitro assays with farnesyl diphosphate as substrate. Compound specific isotope ratio mass spectrometry measurements of the enzyme substrate farnesyl diphosphate (FDP) and the products of all the five cyclases were performed. The calculated δ¹³C value for FDP, based on δ¹³C values and relative amounts of the products, was identical with its measured δ¹³C value, confirming the reliability of the approach and the precision of measurements. The different carbon isotope ratios of the products reflect the complexity of their structure and are correlated with the frequency of carbon–carbon bond forming and breaking steps on their individual biosynthetic pathways. Thus, the analysis of carbon isotopic signatures of terpenes at natural abundance can be used as a powerful tool in elucidation of associated biosynthetic mechanisms of terpene synthases and in future in vivo studies even without ‘touching’ the plant.     

Hemotrophic mycoplasma in Simmental cattle in Bavaria: prevalence,blood parameters,and transplacental transmission of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Mycoplasma haemobos’ and <Emphasis Type="Italic">Mycoplasma wenyonii</Emphasis>

Background

The significance of hemotrophic mycoplasma in cattle remains unclear. Especially in Europe, their epidemiological parameters as well as pathophysiological influence on cows are lacking. The objectives of this study were: (1) to describe the prevalence of ‘Candidatus Mycoplasma haemobos’ (‘C. M. haemobos’) and Mycoplasma wenyonii (M. wenyonii) in Bavaria, Germany; (2) to evaluate their association with several blood parameters; (3) to explore the potential of vertical transmission in Simmental cattle; and (4) to evaluate the accuracy of acridine-orange-stained blood smears compared to real-time polymerase chain reaction (PCR) results to detect hemotrophic mycoplasma. A total of 410 ethylenediaminetetraacetic acid-blood samples from cows from 41 herds were evaluated by hematology, acridine-orange-stained blood smears, and real-time PCR. Additionally, blood samples were taken from dry cows of six dairy farms with positive test results for hemotrophic mycoplasma to investigate vertical transmission of infection.

Results

The period prevalence of both species was 60.24% (247/410), C. M. haemobos 56.59% (232/410), M. wenyonii 8.54% (35/410) and for coinfection 4.88% (20/410). Of the relevant blood parameters, only mean cell volume (MCV), mean cell hemoglobin (MCH), and white blood cell count (WBC) showed differences between the groups of infected and non-infected individuals. There were lower values of MCV (P?<?0.01) and MCH (P?<?0.01) and higher values of WBC (P?<?0.05) in ‘C. M. haemobos’-infected cows. In contrast, co-infected individuals had only higher WBC (P?<?0.05). In M. wenyonii-positive blood samples, MCH was significantly lower (P?<?0.05). Vertical transmission of ‘C. M. haemobos’ was confirmed in two calves. The acridine-orange-method had a low sensitivity (37.39%), specificity (65.97%), positive predictive value (63.70%) and negative predictive value (39.75%) compared to PCR.

Conclusions

‘Candidatus Mycoplasma haemobos’ was more prevalent than M. wenyonii in Bavarian Simmental cattle, but infection had little impact on evaluated blood parameters. Vertical transmission of the infection was rare. Real-time PCR is the preferred diagnostic method compared to the acridine-orange-method.

     

Wettability and Contaminability of Insect Wings as a Function of Their Surface Sculptures

Abstract The wing surfaces of 97 insect species from virtually all relevant major groups were examined by high resolution scanning-electron-microscopy, in order to identify the relationships between the wing microstructures, their wettability with water and their behaviour under the influence of contamination. Isolated wings with contact angles between 31.6° and 155.5° were artificially contaminated with silicate dusts and subsequently fogged until drops of water (“dew”) formed and rolled off. The remaining particles were counted via a digital image analysis system. Remaining particle values between 0.41% and 103% were determined in comparison with unfogged controls. Some insects with very unwettable wings show a highly significant “self-cleaning” effect under the influence of rain or dew. Detailed analysis revealed that there is a correlation between the wettability and the “SM Index” (quotient of wing surface/(body mass)^0.67) with values ranging from 2.42 to 57.0. Furthermore, there is a correlation between the “self-cleaning” effect and the SM Index, meaning that taxa with a high SM Index, e.g. “large-winged” Ephemeroptera, Odonata, Planipennia, and many Lepidoptera, have very unwettable wings and show high particle removal due to dripping water drops. The “small-winged” insects, such as Diptera and Hymenoptera, and insects with elytra, such as Blattariae, Saltatoria, Heteroptera and Coleoptera, show completely opposite effects. This is clearly a result of the fact that species with a high SM Index are, in principle, more restricted in flight by contamination than species with a low SM Index which can also actively clean their own wings. The wings primarily serve a protection function in insects with elytra, so that the effects of contamination are probably of minor importance in these insects. Copyright © 1996 The Royal Swedish Academy of Sciences. Published by Elsevier Science Ltd.     

A putative receptor for Venezuelan equine encephalitis virus from mosquito cells.

We have identified a cellular protein from a continuous mosquito cell line (C6/36) that appears to play a significant role in the attachment of Venezuelan equine encephalitis (VEE) virus to these cells. VEE virus bound to a 32-kDa polypeptide present in the C6/36 plasma membrane fraction, and binding to this polypeptide was dose dependent and saturable and competed with homologous and heterologous alphaviruses. These observations suggest that this polypeptide binds virus via a receptor-ligand interaction. The 32-kDa polypeptide was expressed on the surfaces of C6/36 cells, and monoclonal antibodies directed against either this cell polypeptide or the VEE virus E2 glycoprotein, which is thought to be the viral attachment protein, interfered with virus attachment. Collectively, these data provide evidence suggesting that the 32-kDa polypeptide serves as a receptor for VEE virus infection of cells. We have characterized this cell polypeptide as a laminin-binding protein on the basis of its ability to interact directly with laminin as well as its immunologic cross-reactivity with the high-affinity human laminin receptor.     

High molecular mass glycoproteins associated with the siliceous scales and bristles of Mallomonas splendens (Synurophyceae) may be involved in cell surface development and maintenance

The elaborate scale case of Mallomonas splendens (Synurophyceae) consists of an overlapping arrangement of siliceous scales. In addition, siliceous bristles are attached to specialized base plate scales located at both the anterior and posterior ends of the cells. We have generated monoclonal antibodies against molecules associated with the scale case of M. splendens. One of these antibodies, designated MsS.H9, labelled a proteinaceous epitope of high-molecular-mass cell surface glycoproteins. Immunofluorescence and immunoelectron microscopy demonstrated that only two regions of M. splendens scale cases were labelled by MsS.H9, namely, the upper surface of the scales that contact neighboring scales and the bases of the bristles. Immunoelectron microscopy using thin sections of M. splendens cells showed these labelling sites corresponded to the amorphous material at the sites of scale-to-scale overlap and to a fibrillar complex located at scale-to-bristle attachment sites. Scales and bristles of M. splendens are formed within the cell, in silica deposition vesicles. Immunolabelling of cell sections containing developing scales and bristles showed that MsS.H9 labelling sites were present very early in the formation of these cell surface components. MsS.H9 labelling was also found associated with developing flagellar hairs whereas no labelling was detected on these structures after their deployment onto the flagellum. The location of MsS.H9 labelling sites strongly suggests that the molecule(s) recognized by the antibody plays a role in the adhesion of the individual components making up the scale case of M. splendens.Abbreviations CER chloroplast endoplasmic reticulum - ER endoplasmic reticulum - SDV silica deposition vesicle This work was supported by a grant from the Australian Research Council to R.W. We thank Dr. P. L. Beech for Fig. 13, Dr. L. Perasso for technical assistance and the Plant Cell Biology Group for the use of their monoclonal facilities.     

Pigment-pigment interactions and secondary structure of reconstituted algal chlorophyll a/b-binding light-harvesting complexes of Chlorella fusca with different pigment compositions and pigment-protein stoichiometries

Earlier we have shown by in vitro reconstitution experiments that the pigment composition of the chlorophyll alb-binding light-harvesting complex of the green alga Chlorella fusca could be altered in a relatively broad range (Meyer and Wilhelm 1993). In this study we used these reconstituted complexes of different pigment loading to analyze the excitonic interactions between the pigment molecules and the secondary structure by means of circular dichroism spectra in the visible and the far UV spectral regions, respectively. We found that, in contrast to the expectations, the pigment composition and pigment content hardly affected the circular dichroism spectra in the visible spectral region. Reconstituted complexes, independent of their pigment composition, exhibited the most characteristic circular dichroism bands of the native light-harvesting complex, even if one polypeptide bound only 3 chlorophyll a, 3 chlorophyll b and 1–2 xanthophyll molecules. Full restoration of the protein secondary structure, however, could not be achieved. The -helix content depended significantly on the pigment composition as well as on the pigment-protein ratio of the reconstituted complexes. Further binding of pigments resulted in restoration of the minor excitonic circular dichroism bands, the amplitudes of which depended on the pigment content of the reconstituted complexes. These data suggest that in the reconstitution of light-harvesting complexes a central cluster of pigment molecules plays an important role. Further binding of pigments to the peripheral binding sites appeared also to stabilize the protein secondary structure of the reconstituted complexes.Abbreviations CD- circular dichroism - LHC- chlorophyll a/b light-harvesting complex(es) - LHC II- light-harvesting complex(es) of Photosystem II of higher plants - LHCP- light-harvesting Chl a/b-binding protein(s) - PP- polypeptide(s)     

A Long-Chain Secondary Alcohol Dehydrogenase from Rhodococcus erythropolis ATCC 4277

A NAD-dependent secondary alcohol dehydrogenase has been purified from the alkane-degrading bacterium, Rhodococcus erythropolis ATCC 4277. The enzyme was found to be active against a broad range of substrates, particularly long-chain secondary aliphatic alcohols. Although optimal activity was observed with linear 2-alcohols containing between 6 and 11 carbon atoms, secondary alcohols as long as 2-tetradecanol were oxidized at 25% of the rate seen with mid-range alcohols. The purified enzyme was specific for the S-(+) stereoisomer of 2-octanol and had a specific activity for 2-octanol of over 200 (mu)mol/min/mg of protein at pH 9 and 37(deg)C, 25-fold higher than that of any previously reported S-(+) secondary alcohol dehydrogenase. Linear primary alcohols containing between 3 and 13 carbon atoms were utilized 20- to 40-fold less efficiently than the corresponding secondary alcohols. The apparent K(infm) value for NAD(sup+) with 2-octanol as the substrate was 260 (mu)M, whereas the apparent K(infm) values for the 2-alcohols ranged from over 5 mM for 2-pentanol to less than 2 (mu)M for 2-tetradecanol. The enzyme showed moderate thermostability (half-life of 4 h at 60(deg)C) and could potentially be useful for the synthesis of optically pure stereoisomers of secondary alcohols.