Lysophosphatidic acid receptor 1 inhibitor,AM095, attenuates diabetic nephropathy in mice by downregulation of TLR4/NF-κB signaling and NADPH oxidase

Diabetic nephropathy (DN) is one of the major long-term complications of diabetes. Lysophosphatidic acid (LPA) signaling has been implicated in renal fibrosis. In our previous study, we found that the LPA receptor 1/3 (LPAR1/3) antagonist, ki16425, protected against DN in diabetic db/db mice. Here, we investigated the effects of a specific pharmacological inhibitor of LPA receptor 1 (LPA1), AM095, on DN in streptozotocin (STZ)-induced diabetic mice to exclude a possible contribution of LPAR3 inhibition. AM095 treatment significantly reduced albuminuria and the albumin to creatinine ratio and significantly decreased the glomerular volume and tuft area in the treated group compared with the STZ-vehicle group. In the kidney of STZ-induced diabetic mice, the expression of LPAR1 mRNA and protein was positively correlated with oxidative stress. AM095 treatment inhibited LPA-induced reactive oxygen species production and NADPH oxidase expression as well as LPA-induced toll like receptor 4 (TLR4) expression in mesangial cells and in the kidney of STZ-induced diabetic mice. In addition, AM095 treatment suppressed LPA-induced pro-inflammatory cytokines and fibrotic factors expression through downregulation of phosphorylated NFκBp65 and c-Jun N-terminal kinases (JNK) in vitro and in the kidney of STZ-induced diabetic mice. Pharmacological or siRNA inhibition of TLR4 and NADPH oxidase mimicked the effects of AM095 in vitro. In conclusion, AM095 is effective in preventing the pathogenesis of DN by inhibiting TLR4/NF-κB and the NADPH oxidase system, consequently inhibiting the inflammatory signaling cascade in renal tissue of diabetic mice, suggesting that LPAR1 antagonism might provide a potential therapeutic target for DN.     

Flow cytometric method for quantifying viable Mycoplasma agassizii,an agent of upper respiratory tract disease in the desert tortoise (Gopherus agassizii)

Aims: Mycoplasma agassizii can cause upper respiratory tract disease in the threatened desert tortoise of the Southwestern United States. Two technical challenges have impeded critical microbiological studies of this microorganism: (i) its small size limits the use of light microscopy for cell counting and (ii) its extremely slow growth in broth and agar cultures impedes colony counting. Our aim was to develop a rapid and sensitive flow cytometric method using a vital fluorescent dye to enumerate viable M. agassizii cells. Methods and Results: Here, we demonstrate that the nonfluorescent molecule 5‐carboxyfluorescein (5‐CF) diacetate acetoxymethyl ester penetrates M. agassizii cell membranes and it is converted in the cytoplasm to the fluorescent molecule 5‐CF by the action of intracellular esterases. Labelled mycoplasma cells can be easily detected by flow cytometry, and cultures with as few as 100 viable mycoplasma cells ml^?1 can be labelled and counted in less than 1 h. Experiments using temperature‐induced cell death demonstrated that only viable M. agassizii cells are labelled with this procedure. Conclusions: A rapid and sensitive flow cytometric technique has been developed for enumerating viable M. agassizii cells. Significance and Impact of the Study: This technique should facilitate basic immunological, biochemical and pharmacological studies of this important pathogen which may lead to new diagnostic and therapeutic methods.     

Spore ontogeny of the arbuscular mycorrhizal fungus<Emphasis Type="Italic"> Archaeospora trappei</Emphasis> (Ames &; Linderman) Morton &; Redecker (<Emphasis Type="Italic">Archaeosporaceae</Emphasis>)

Arbuscular mycorrhizal (AM) fungi in the genus Archaeospora (family Archaeosporaceae) contain both monomorphic and dimorphic species. The synanamorphism is often hard to discern without ontogenetic observations. Here, the spore ontogeny of Ar. trappei is reported from single species pot culture studies. The sporogenous hypha swelled up to a terminal sporiferous saccule and produced a lateral spore primordium on its neck. The saccule expanded fully before the spore primordium emerged. The saccule transferred its contents into the expanding spore and collapsed while wall differentiation continued inside the spore. The spore wall of Ar. trappei differentiated sequentially, in discrete steps, as in Acaulosporaceae members. In contrast, Ar. trappei produced a simplified spore wall in which the components differed in chemical and physical characteristics from those of the Acaulosporaceae members. Ontogenetic studies confirmed Ar. trappei to be monomorphic and producing acaulosporoid spores. The fungus is a new record to New Zealand.     

Interval-integration underlies amplitude modulation band-suppression selectivity in the anuran midbrain

We examined the mechanisms that underlie band-suppression amplitude modulation selectivity in the auditory midbrain of anurans. Band-suppression neurons respond well to low (5–10 Hz) and high (>70 Hz) rates of sinusoidal amplitude modulation, but poorly, if at all, to intermediate rates. The effectiveness of slow rates of sinusoidal amplitude modulation is due to the long duration of individual pulses; short-duration pulses (<10 ms) failed to elicit spikes when presented at 5–10 pulses s^–1. Each unit responded only after a threshold number of pulses (median=3, range=2–5) were delivered at an optimal rate. The salient stimulus feature was the number of consecutive interpulse intervals that were within a cell-specific tolerance. This interval-integrating process could be reset by a single long interval, even if preceded by a suprathreshold number of intervals. These findings indicate that band-suppression units are a subset of interval-integrating neurons. Band-suppression neurons differed from band-pass interval-integrating cells in having lower interval-number thresholds and broader interval tolerance. We suggest that these properties increase the probability of a postsynaptic spike, given a particular temporal pattern of afferent action potentials in response to long-duration pulses, i.e., predispose them to respond to slow rates of amplitude modulation. Modeling evidence is provided that supports this conclusion.Abbreviations AM amplitude modulation - PRR pulse repetition rate - SAM sinusoidal amplitude modulation     

Effectiveness of recruitment behavior in stingless bees (Apidae,Meliponini)

Summary We examined the ability of stingless bees to recruit nest mates to a food source (i) in group foraging species laying pheromone trails from the food to the nest (Trigona recursa Smith, T. hypogea Silvestri, Scaptotrigona depilis Moure), (ii) in solitary foraging species with possible but still doubtful communication of food location inside the nest (Melipona seminigra Friese, M. favosa orbignyi Guérin), and (iii) in species with a less precise (Nannotrigona testaceicornis Lep., Tetragona clavipes Fab.) or no communication (Frieseomelitta varia Lep.). The bees were allowed to collect food (sugar solution or liver in the necrophageous species) ad libitum and the forager number to accumulate, as it would do under normal unrestrained conditions. The median number of bees collecting differed considerably among the species (1.0–1436.5). It was highest in the species employing scent trails. The time course of recruitment was characteristic for most of the species and largely independent of the number of foragers involved. The two Melipona species recruited other bees significantly faster than T. recursa, S. depilis, and N. testaceicornis during the first 10 to 30 minutes of an experiment. In species laying a scent trail to guide nestmates to a food source the first recruits appeared with a delay of several minutes followed by a quick increase in forager number. The median time required to recruit all foragers available differed among the species between 95.0 and 240.0 min. These differences can at least partly be explained by differences in the recruitment mechanisms and do not simply follow from differences in colony biomass.     

Molecular diversity of arbuscular mycorrhizal fungi and patterns of host association over time and space in a tropical forest

We have used molecular techniques to investigate the diversity and distribution of the arbuscular mycorrhizal (AM) fungi colonizing tree seedling roots in the tropical forest on Barro Colorado Island (BCI), Republic of Panama. In the first year, we sampled newly emergent seedlings of the understory treelet Faramea occidentalis and the canopy emergent Tetragastris panamensis, from mixed seedling carpets at each of two sites. The following year we sampled surviving seedlings from these cohorts. The roots of 48 plants were analysed using AM fungal-specific primers to amplify and clone partial small subunit (SSU) ribosomal RNA gene sequences. Over 1300 clones were screened for random fragment length polymorphism (RFLP) variation and 7% of these were sequenced. Compared with AM fungal communities sampled from temperate habitats using the same method, the overall diversity was high, with a total of 30 AM fungal types identified. Seventeen of these types have not been recorded previously, with the remainder being similar to types reported from temperate habitats. The tropical mycorrhizal population showed significant spatial heterogeneity and nonrandom associations with the different hosts. Moreover there was a strong shift in the mycorrhizal communities over time. AM fungal types that were dominant in the newly germinated seedlings were almost entirely replaced by previously rare types in the surviving seedlings the following year. The high diversity and huge variation detected across time points, sites and hosts, implies that the AM fungal types are ecologically distinct and thus may have the potential to influence recruitment and host composition in tropical forests.     

Autogenic training: a meta-analysis of clinical outcome studies

Autogenic training (AT) is a self-relaxation procedure by which a psychophysiological determined relaxation response is elicited. A meta-analysis was performed to evaluate the clinical effectiveness of AT. Seventy-three controlled outcome studies were found (published 1952–99). Sixty studies (35 randomized controlled trials [RCT]) qualified for inclusion in the meta-analysis. Medium-to-large effect sizes (ES) occurred for pre–post comparisons of disease-specific AT-effects, with the RCTs showing larger ES. When AT was compared to real control conditions, medium ES were found. Comparisons of AT versus other psychological treatment mostly resulted in no effects or small negative ES. This pattern of results was stable at follow-up. Unspecific AT-effects (i.e., effects on mood, cognitive performance, quality of life, and physiological variables) tended to be even larger than main effects. Separate meta-analyses for different disorders revealed a significant reduction of the heterogeneity of ES. Positive effects (medium range) of AT and of AT versus control in the meta-analysis of at least 3 studies were found for tension headache/migraine, mild-to-moderate essential hypertension, coronary heart disease, asthma bronchiale, somatoform pain disorder (unspecified type), Raynaud's disease, anxiety disorders, mild-to-moderate depression/dysthymia, and functional sleep disorders.     

AM1* parameters for phosphorus,sulfur and chlorine

An extension of the AM1 semiempirical molecular orbital technique, AM1*, is introduced. AM1* uses AM1 parameters and theory unchanged for the elements H, C, N, O and F. The elements P, S and Cl have been reparameterized using an additional set of d orbitals in the basis set and with two-center core–core parameters, rather than the Gaussian functions used to modify the core–core potential in AM1. Voityuk and Röschs AM1(d) parameters have been adopted unchanged for AM1* with the exception that new core–core parameters are defined for Mo–P, Mo–S and Mo–Cl interactions. Thus, AM1* gives identical results to AM1 for compounds with only H, C, N, O, and F, AM1(d) for compounds containing Mo, H, C, N, O and F only, but differs for molybdenum compounds containing P, S or Cl. The performance and typical errors of AM1* are discussed.Electronic Supplementary Material Supplementary material is available in the online version of this article at . Tables 2 and 4–7 and a full list (Tables S1, S2) of geometrical parameters and barrier heights are given in the supplementary material.This revised version was published online in September 2003.     

我国东、南沿海地区AM真菌群落生态分布研究

在我国东、南沿海地区AM真菌资源调查的基础上研究了AM真菌的群落生态分布和特征。证明球囊霉属在该地区的出现频度和相对多度都最高,其次是硬囊霉属。该区内南四省AM真菌丰度、密度和物种多样性指数均比北三省高。聚类分析结果和上述相似,说明我国东、南沿海各省AM真菌的丰度、频度、密度、多度和多样性特征由北向南呈规律性变化。     

国家重点保护野生植物的保护现状及潜在分布区预测分析

野生植物是自然生态系统的重要组成部分,中国是野生植物种类最丰富的国家之一。研究国家重点保护野生植物的分布特征、保护现状以及潜在分布区,对于制定与支持生物多样性保护策略具有重要意义。该研究基于1 032种(隶属于129科315属)国家重点保护野生植物,利用前5%丰富度算法识别其热点地区,并与自然保护区叠加评估其保护成效、确定保护空缺,进而运用MaxEnt模型预测了国家重点保护野生植物的潜在分布区分布与变化趋势。结果表明:(1)中国南部和西南部是国家重点保护野生植物物种丰富度最高的地区,尤其是四川中部、云南南部和东南部、广西北部、广东北部与海南。(2)热点网格的保护成效分析表明,171个(85.50%)热点网格得到了有效保护(含80.50%的物种),29个(14.50%)热点网格未得到自然保护区的保护(含51.20%物种)。(3)通过比较当前与未来气候变化下国家重点保护野生植物的潜在分布区分布,发现未来潜在分布区将向西藏东南部、广西西南部、广东南部以及福建南部等地扩张,而向环四川盆地、云南南部和贵州南部等地缩减。因此,需要加强这些区域生物多样性的动态监测,持续关注气候变化对该区域国家重点保护野生植物的影响。基于该研究所确定的热点网格、保护成效以及潜在分布区的分析结果,可为国家重点保护野生植物多样性优先保护区的确定和保护政策的制定提供有力的数据支持与参考。