Disparate determinants of summer and winter diet selection of a generalist herbivore,Ochotona princeps

The North American pika, Ochotona princeps, is a generalist herbivore that simultaneously selects two distinct diets: one consumed immediately (summer diet), the other harvested, transported, and stored for later consumption (winter diet). I investigated factors influencing diet selection at two sites on the West Knoll of Niwot Ridge, Boulder County, Colorado during 1991 and 1992. The composition of summer and winter diets differed significantly from each other as well as from the relative abundance of food items in the environment. Thus, pikas were not foraging randomly for either diet. To explore winter and summer diet selection, I tested two existing hypotheses: (1) that plant morphology restricts the winter diet breadth to plants that are easily harvested and large, and thereby maximizes the amount collected per foraging effort, or (2) to compensate for nutrients lost during storage, pikas bias their winter diet with high-nutrient species. I also tested the hypothesis that plant secondary compounds may be higher in the winter diet either because they function as preservatives or because pikas delay consumption of these species until the toxins degrade. For individual dietary items, I measured energy, nitrogen, water, fiber, total phenolic, condensed tannin, and astringency contents. There was little evidence to suggest that morphology excluded plants from the winter diet. Plant size was not a good predictor of abundance in the winter diet. Even after harvesting costs had been experimentally removed, cushion plants were still not included in the winter diet. There was weak support for an effect of nutrients on winter diet selection; in three of four cases, the winter diet was significantly lower in water and higher in total energy content as predicted by the nutrient compensation hypothesis. However, other nutrients exhibited no consistent pattern. Nutrients were not reliable predictors of the winter diet in multiple regression analyses. There was strong support for the hypothesis of manipulation of secondary compounds. The winter diet was significantly higher in total phenolics and astringency. Total phenolics were consistent predictors of the winter diet in multiple regression analyses. The winter diets of six additional pika populations contained plant species high in secondary compounds. The results suggest that pikas preferentially select plants with high levels of secondary compounds for their winter diet, possibly because the presence of such compounds promotes preservation of the cache. This behavior may also enable the exploitation of an otherwise unusable food resource, i.e., toxic plants.     

FFAR4 regulates cardiac oxylipin balance to promote inflammation resolution in HFpEF secondary to metabolic syndrome

Heart failure with preserved ejection fraction (HFpEF) is a complex clinical syndrome, but a predominant subset of HFpEF patients has metabolic syndrome (MetS). Mechanistically, systemic, nonresolving inflammation associated with MetS might drive HFpEF remodeling. Free fatty acid receptor 4 (Ffar4) is a GPCR for long-chain fatty acids that attenuates metabolic dysfunction and resolves inflammation. Therefore, we hypothesized that Ffar4 would attenuate remodeling in HFpEF secondary to MetS (HFpEF-MetS). To test this hypothesis, mice with systemic deletion of Ffar4 (Ffar4KO) were fed a high-fat/high-sucrose diet with L-NAME in their water to induce HFpEF-MetS. In male Ffar4KO mice, this HFpEF-MetS diet induced similar metabolic deficits but worsened diastolic function and microvascular rarefaction relative to WT mice. Conversely, in female Ffar4KO mice, the diet produced greater obesity but no worsened ventricular remodeling relative to WT mice. In Ffar4KO males, MetS altered the balance of inflammatory oxylipins systemically in HDL and in the heart, decreasing the eicosapentaenoic acid-derived, proresolving oxylipin 18-hydroxyeicosapentaenoic acid (18-HEPE), while increasing the arachidonic acid-derived, proinflammatory oxylipin 12-hydroxyeicosatetraenoic acid (12-HETE). This increased 12-HETE/18-HEPE ratio reflected a more proinflammatory state both systemically and in the heart in male Ffar4KO mice and was associated with increased macrophage numbers in the heart, which in turn correlated with worsened ventricular remodeling. In summary, our data suggest that Ffar4 controls the proinflammatory/proresolving oxylipin balance systemically and in the heart to resolve inflammation and attenuate HFpEF remodeling.     

Toxicity of aldehyde products of lipid peroxidation to adult Schistosoma intercalatum in vitro

The host's immune response results in oxidative damage to parasite membranes. Known aldehyde breakdown products from lipid peroxidation have been investigated for their in vitro toxicity to Schistosoma intercalatum. Saturated and monounsaturated aldehydes were found to be relatively non-toxic, whilst dienal and hydroxyenal aldehydes had LD₅₀ values in the range of 10–20 μM. Conversion of the toxic aldehydes to their corresponding alcohols or glutathione conjugates reduced toxicity to S. intercalatum by one or two orders of magnitude. This suggests that parasite detoxification enzymes might be useful targets for chemotherapy and raises the possibility of combining chemo- and immunotherapy.     

Bioactive metabolites from the endophytic fungus Ampelomyces sp. isolated from the medicinal plant Urospermum picroides

Extracts of cultures grown in liquid or on solid rice media of the fungal endophyte Ampelomyces sp. isolated from the medicinal plant Urospermum picroides exhibited considerable cytotoxic activity when tested in vitro against L5178Y cells. Chromatographic separation yielded 14 natural products that were unequivocally identified based on their ¹H and ¹³C NMR as well as mass spectra and comparison with previously published data. Six compounds (2, 4, 5, 7, 9 and 11) were natural products. Both fungal extracts differed considerably in their secondary metabolites. The extract obtained from liquid cultures afforded a pyrone (2) and sulfated anthraquinones (7 and 9) along with the known compounds 1, 3, 6 and 8. When grown on solid rice medium the fungus yielded three compounds 4, 5 and 11 in addition to several known metabolites including 6, 8, 10, 12, 13 and 14. Compounds 4, 8 and 10 showed the strongest cytotoxic activity against L5178Y cells with EC₅₀ values ranging from 0.2–7.3 μg/ml. Furthermore, 8 and 10 displayed antimicrobial activity against the Gram-positive pathogens, Staphylococcus aureus, S. epidermidis and Enterococcus faecalis at minimal inhibitory concentrations (MIC) of 12.5 μg/ml and 12.5–25 μg/ml, respectively. Interestingly, 6 and 8 were also identified as constituents of an extract derived from a healthy plant sample of the host plant U. picroides thereby indicating that the production of bioactive natural products by the endophyte proceeds also under in situ conditions within the host plant.     

The ovarian phenotype of the aromatase knockout (ArKO) mouse

Targeted disruption of exon 9 of the cyp19 gene gives rise to a non-functional aromatase enzyme incapable of converting androgens to oestrogens. The aromatase knockout (ArKO) mouse is, thus, characterised by a dysfunctional pituitary-gonadal axis, which manifests in non-detectable levels of oestrogen in serum. These mice also exhibit elevated levels of circulating gonadotrophins (luteinising hormone (LH) and follicle stimulating hormone (FSH)) and testosterone. The ArKO mouse is infertile due to folliculogenic disruption and a failure to ovulate. The age-dependent ovarian phenotype revealed a block in follicular development at the antral stage and a complete absence of corpora lutea. By 21–23 weeks of age haemorrhagic cystic follicles were present and by 1 year there were abnormal follicles, an absence of secondary and antral follicles and atretic primary follicles. Interstitial tissue remodelling was extensive and exemplified by an increase in collagen deposition and an influx of macrophages, coincident with the loss of follicles. In mice, maintained on a soy-free and, thus, phytoestrogen-free diet, the ovarian phenotype was accelerated and exacerbated. In conclusion, the ovarian phenotype of the ArKO mouse can be attributed to the altered hormonal environment brought about by the absence of aromatase and the failure of androgens to be converted to oestrogens in the presence of elevated gonadotropins.     

The Role of Anorexia in Resistance and Tolerance to Infections in Drosophila

Most infections induce anorexia but its function, if any, remains unclear. Because this response is common among animals, we hypothesized that infection-induced diet restriction might be an adaptive trait that modulates the host's ability to fight infection. Two defense strategies protect hosts against infections: resistance, which is the ability to control pathogen levels, and tolerance, which helps the host endure infection-induced pathology. Here we show that infected fruit flies become anorexic and that diet restriction alters defenses, increasing the fly's tolerance to Salmonella typhimurium infections while decreasing resistance to Listeria monocytogenes. This suggests that attempts to extend lifespan through diet restriction or the manipulation of pathways mimicking this process will have complicated effects on a host's ability to fight infections.     

地衣型真菌石果衣(疣孢菌科，子囊菌门)次级代谢产物的多样性

地衣能够产生大量的新型次级代谢产物,但以往对石果衣Endocarpon pusillum的研究中未能检测出任何次级代谢产物。然而,对其共生菌进行基因组测序发现其中含有14个沉默的PKS基因和2个沉默的NRPS基因。在此研究中,为激活其途径,使用了优化马铃薯培养基和大米培养基对石果衣共生真菌进行了培养。从优化马铃薯培养物中分离得到9个次级代谢产物,包括2个新的异吲哚-1-酮类化合物(1, 2)。而从大米培养物中分离到3个已知化合物和1个新的萘醌类化合物(9)。通过核磁共振和质谱数据确定了新化合物的结构。研究结果表明,大量地衣中未能检测出任何次级代谢产物,或仅能检测出少量次级代谢产物,可能与其基因组中的沉默基因有关。因此,通过对沉默基因的激活方法为地衣次级代谢产物资源的研究与开发开辟了有效途径。     

Chemistry of Cirsium and Carduus: a role in ecological risk assessment for biological control of weeds?

Prediction of host plant range and ecological impact of exotic phytophagous insects, such as insects for classical biological control of weeds, represents a major challenge. Recently, the flowerhead weevil (Rhinocyllus conicus Fröl.), introduced from Europe into North America to control exotic thistles (Carduus spp.), has become invasive. It feeds heavily on some, but not all species of native North American thistles (Cirsium spp.). We hypothesized that such non-target use among native plants could be better predicted by knowledge of characteristic chemical profiles of secondary compounds to supplement the results of host specificity testing. To evaluate this hypothesis, we reviewed the literature on the chemistry of Cirsium and Carduus thistles. We asked what compounds are known to be present, what is known about their biological activity, and whether such information on chemical profiles would have better predicted realized host range and ecological effects of R. conicus in North America. We found an extensive, but incomplete literature on the chemistry of true thistles. Two main patterns emerged. First, consistent chemical similarities and interesting differences occur among species of thistles. Second, variation occurs in biologically active groups of characteristic compounds, specifically flavonoids, sterols, alkaloids and phenolic acids, that are known to influence host plant acceptance, selection, and feeding by phytophagous insects. Surprisingly, sesquiterpene lactones, which are characterisitic in closely related Asteraceae, have not been extensively reported for Cirsium or Carduus. The minimal evidence on sesquiterpene lactones may reflect extraction methods vs. true absence. In summary, our review suggests further research on thistle chemistry in insect feeding is warranted. Also, since the exotic Canada thistle (Cirsium arvense) is an invasive thistle of current concern in North America, such research on mechanisms underlying host range expansion by exotic insects would be useful.     

Plant Secondary Compounds as Diuretics: An Overlooked Consequence

Plant secondary compounds are deterrents and toxins to a varietyof herbivores. The effect of secondary compounds on water balanceof herbivores is virtually unexplored, yet many secondary compoundsare renowned for their diuretic effects in humans and laboratoryrats. We review data from the ethnopharmocological literatureon plants with diuretic effects. We also present our data fromexperiments on water intake of specialist (Neotoma stephensi)and generalist woodrats (N. albigula) consuming plant secondarycompounds from their natural diet. We measured effects of dietarysecondary compounds on voluntary water consumption, urine volumeand urine osmolarity. Ingestion of secondary compounds increasedwater intake and urine output and decreased urine osmolarityin both species. However, the generalist was more impacted bydietary secondary compounds than the specialist. Our resultscombined with that from the literature suggest that diuresismay be a prevalent consequence of ingestion of secondary compounds.Many herbivores live in arid habitats with limited access tofree-standing water, thus an increase in the desire for watermay have profound consequences on foraging behavior and fitness.     

Antihyperglycemic activity of compounds isolated from Indian medicinal plants

Eleven antidiabetic Indian medicinal plants were investigated in streptozotocin induced diabetic rat model and provided scientific validation to prove their antihyperglycemic activity. Antidiabetic principles from five plants were isolated. All the compounds isolated were evaluated for antihyperglycemic activity in streptozotocin induced diabetic rat model and activities were compared with standard drug metformin. Some compounds were also screened in db/db mice. Two compounds (PP-1 and PP-2) inhibited significantly the activity of PTPase-1B in an in vitro system. This might be the underlying mechanism of antihyperglycemic activity of these compounds.     

SALIVARY ANDROGEN-BINDING PROTEIN (ABP) MEDIATES SEXUAL ISOLATION IN MUS MUSCULUS

We wanted to determine whether the microevolution of the mouse salivary androgen-binding protein (ABP) Alpha subunit gene (Abpa) could mediate sexual selection and thereby have a potential role in maintaining gene pool integrity where radiating mouse subspecies make secondary contact. This hypothesis is based upon previous work in this laboratory, which has shown that each subspecies apparently has its own allele and that these alleles have a 25-fold excess of nonsynonymous/synonymous base substitutions compared to an average protein under purifying selection. We provide direct evidence for ABP-assortative mate selection in a laboratory setting: Mus musculus domesticus and M. m. musculus female mice recognize and discriminate between the territories of male mice that essentially differ solely in their Abpa genotype and, when the males are present, the female prefers to mate with the one of her own ABP type. The observation that females could differentiate between the territories of the two males when those mice were absent suggests that the males marked their territories with ABP. In this study, we also detected ABP on the pelts of male mice and in their environment. It is likely that the animals apply the protein to their pelts by licking and that it is then deposited in their surroundings. We suggest that females of the two subspecies are able to discriminate between males of those subspecies on the basis of this protein molecule. Mouse salivary ABP might present a worthwhile system with which to study a prezygotic isolation mechanism in a mammal.     

Anti-inflammatory activities of triterpenoid saponins from Polygala japonica

Bioassay-guided investigation was performed to identify the active constituents from a methanol extract of Polygala japonica, a folk medicinal plant widely used in China to treat inflammatory diseases. The n-BuOH and EtOAc fractions of the P. japonica methanol extract, which show significant anti-inflammatory activity in in vivo test, were further subjected to column chromatography to afford six triterpene glycosides, marked here as saponins 1–6. All compounds were evaluated for their anti-inflammatory activity in the carageenan-induced mouse paw edema test, and saponins 1, 4 and 5 showed significantly anti-inflammatory effects on both phases of carageenan-induced acute paw edema in mice. Saponin 5 was also found to significantly inhibit the production of inflammatory mediators – nitric oxide (NO) in LPS-stimulated RAW264.7 macrophages, with no obvious effects on macrophage viability.     

新疆药用植物天山雪莲及红景天内生真菌的分离与初步鉴定

本文采用常规组织块分离法及直接克隆测序的方法初步鉴定了新疆天山地区代表性的两种药用植物天山雪莲和红景天根内可培养及不可培养的内生真菌。从两种药用植物根中共分离获得34株内生真菌,除7株鉴定为镰孢菌属Fusarium外,其余菌株均具有暗色、有隔菌丝,不产生孢子;从形态上看,属典型的深色有隔内生真菌（dark septate endophyte,DSE）。采用直接克隆测序的方法,从两种植物的根中共获得143个真菌克隆子。依据核糖体内转录间隔区（internal transcribed spacer,ITS）序列鉴定多数克隆子归属于柔膜菌目Helotiales及格孢腔菌目Pleosporales,与已知的DSE如背芽突霉属Cadophora、瓶头霉属Phialocephala、瓶霉属Phialophora以及Leptodontidium等有90%以上的核苷酸序列相似性。该结果显示DSE在天山雪莲和红景天根部内生真菌中占优势,暗示其可能参与了宿主植物对高寒、强辐射环境的生态适应。     

Atamestane: An aromatase inhibitor for the treatment of benign prostatic hyperplasia. A short review

Benign prostatic hyperplasia (BPH) is the most common neoplastic growth in men and is the most frequent cause of urinary flow obstruction at the bladder neck. In addition to the clear evidence in favor of the androgen dependency of BPH, the involvement of the stroma, stromal-epithelial interaction and the role of estrogens have gained much interest in connection with the pathogenesis of this disease. For this reason, specific aromatase inhibitors such as atamestane (1-methyl-1,4-androstadiene-3,17-dione) have recently attracted attention due to their potential use in the treatment of BPH. The pharmacological action of atamestane as a new competitive and irreversible inhibitor of estrogen biosynthesis has been evaluated in mice, rats, rabbits, dogs, monkeys and in man. In all species tested so far, atamestane lacks other intrinsic hormonal or antihormonal activities and shows no inhibition of other cytochrome-P₄₅₀ dependent enzymes of adrenal steroidogenesis. However, it inhibits the estrogen-related negative feed-back. The extent and consequence of the induced counter-regulation of the pituitary-hypothalamic axis show major sex- and species-specific differences. In BPH animal models, atamestane is highly effective in inhibiting estrogen-induced hyperplastic changes in the fibromuscular stroma of the prostate in androstenedione-treated dogs and monkeys. In male volunteers and BPH patients, atamestane induces an expected dose-dependent reduction of serum estrogen concentrations with slight increases in androgen level. In conclusion, all available results indicate that atamestane is a selective (no inhibition of adrenal function), pure (= specific—no endocrine side-effects) and highly effective steroidal aromatase inhibitor with excellent safety profile. Based on our preliminary results aromatase inhibitors seem to be promising compounds for the treatment of BPH.     

基于广泛靶向代谢组学的竹黄活性成分分析

竹黄是我国一种重要的药用真菌,在医学、农业、食品等方面应用广泛且前景可观。为深入挖掘竹黄中有药理活性的有效化学成分,了解其在生长发育过程中不同时期代谢物的变化规律,利用广泛靶向代谢组学技术检测了竹黄子座不同发育时期的代谢物,找出差异代谢物并进行代谢通路分析。从竹黄子座中共检测出612种代谢物,前期和中期特有27种代谢物。黄酮类、奎宁酸、香豆素等具有良好生物活性的化合物首次在竹黄中被检测到。筛选出的差异代谢物主要是脂质、氨基酸、核苷酸、黄酮类、萜类、有机酸等物质,其中黄酮和氨基酸类化合物占主要地位。通过对代谢通路富集分析,获得6条具有显著意义的代谢途径。黄酮类化合物被认为是除竹红菌素外与竹黄药效有重要联系的化合物。本研究为竹黄药用机理及有效成分深入研究提供了一定的理论基础,为竹黄有效成分的代谢途径解析提供参考。     

Extended Exenatide Administration Enhances Lipid Metabolism and Exacerbates Pancreatic Injury in Mice on a High Fat,High Carbohydrate Diet

This study expanded upon a previous study in mice reporting a link between exenatide treatment and exocrine pancreatic injury by demonstrating temporal and dose responses and providing an initial mechanistic hypothesis. The design of the present study included varying lengths of exenatide exposure (3, 6 weeks to 12 weeks) at multiple concentrations (3, 10, or 30 µg/kg) with multiple endpoints (histopathology evaluations, immunoassay for cytokines, immunostaining of the pancreas, serum chemistries and measurement of trypsin, amylase, and, lipase, and gene expression profiles). Time- and dose-dependent exocrine pancreatic injury was observed in mice on a high fat diet treated with exenatide. The morphological changes identified in the pancreas involved acinar cell injury and death (autophagy, apoptosis, necrosis, and atrophy), cell adaptations (hypertrophy and hyperplasia), and cell survival (proliferation/regeneration) accompanied by varying degrees of inflammatory response leading to secondary injury in pancreatic blood vessels, ducts, and adipose tissues. Gene expression profiles indicated increased signaling for cell survival and altered lipid metabolism in exenatide treated mice. Immunohistochemistry supported gene expression findings that exenatide caused and/or exacerbated pancreatic injury in a high fat diet environment potentially by further increasing high fat diet exacerbated lipid metabolism and resulting oxidative stress. Further investigation is required to confirm these findings and determine their relevance to human disease.     

Metabolic engineering of plant secondary products

Plants interact with their environment by producing a diverse array of secondary metabolites. Many of these compounds are valued for their medicinal, industrial or agricultural properties. Other secondary products are toxic or otherwise undesirable and can reduce the commercial value of crops. Gene transfer technology offers new opportunities to modify directly plant secondary product synthesis through metabolic engineering. This article reviews some of the strategies which have been used to increase or decrease the synthesis of specific plant metabolites, as well as methods for expanding the biosynthetic capabilities of individual species.     

Probiotics function mechanistically as delivery vehicles for neuroactive compounds: Microbial endocrinology in the design and use of probiotics

I hypothesize here that the ability of probiotics to synthesize neuroactive compounds provides a unifying microbial endocrinology-based mechanism to explain the hitherto incompletely understood action of commensal microbiota that affect the host's gastrointestinal and psychological health. Once ingested, probiotics enter an interactive environment encompassing microbiological, immunological, and neurophysiological components. By utilizing a trans-disciplinary framework known as microbial endocrinology, mechanisms that would otherwise not be considered become apparent since any candidate would need to be shared among all three components. The range of neurochemicals produced by probiotics includes neurochemicals for which receptor-based targets on immune and neuronal elements (intestinal and extra-intestinal) have been well characterized. Production of neurochemicals by probiotics therefore allows for their consideration as delivery vehicles for neuroactive compounds. This unifying microbial endocrinology-based hypothesis, which may facilitate the selection and design of probiotics for clinical use, also highlights the largely unrecognized role of neuroscience in understanding how microbes may influence health.     

灵芝β-葡萄糖苷酶基因的克隆与功能分析

灵芝是我国著名的药用真菌,具有抗癌、抗肿瘤等功效。灵芝酸属于三萜类化合物,是灵芝的主要活性成分,并已成为评价灵芝品质的重要指标之一。β-葡萄糖苷酶(β-glucosidase,BG)是次生代谢产物合成途径中的关键限速酶,能够调节次生代谢产物的生物合成。本研究通过同源序列比对,注释获得了灵芝β-葡萄糖苷酶基因(GlBG),并通过RNAi技术对灵芝β-葡萄糖苷酶进行功能分析。序列分析结果显示GlBG基因的DNA全长为2 759 bp,包含7个外显子和6个内含子,编码793个氨基酸,其编码的蛋白序列中含有β-糖苷水解酶的2个保守结构域。灵芝β-葡萄糖苷酶基因的沉默转化子中灵芝酸含量比野生型菌株的灵芝酸含量平均降低了38%,并且灵芝酸生物合成途径中的关键酶基因(hmgs、hmgr、fps、sqs和osc)的表达量也显著下降,实验结果表明灵芝β-葡萄糖苷酶在灵芝酸生物合成过程中具有重要作用,并为灵芝次生代谢途径及其调控机制提供了参考。