Selected culturable enteric bacterial populations are modified by diet acidification and the growth promotant Tylosin

AIMS: To determine the effect of diet acidification and an in-feed antibiotic growth promotant (Tylosin, Ty) on selected culturable bacterial populations in the gastrointestinal tract (GIT) of mice. METHODS AND RESULTS: Female C57Bl mice were given a standard diet supplemented with Acid Pak (AP) or Ty in the drinking water. After 21 days, lumen and adherent populations of Enterobacteriaceae, enterococci/streptococci, and lactic acid bacteria (LAB) from the ileum, caecum, colon and faeces were enumerated. General intestinal health was assessed by the frequency of haemolytic bacteria in the different intestinal compartments. Contrary to expectations, AP and Ty significantly increased haemolytic bacteria in the lumen of the caecum and colon (P<0.05). The small but significant growth-enhancing effect of Ty (P<0.05) was associated with decreases in enterococci/streptococci and surprisingly, LAB, as well as increases in coliforms. AP, which failed to improve growth rates, reduced coliforms, had limited effects on enterococci/streptococci, and specifically failed to promote the growth of LAB populations in all intestinal compartments. Ty supplementation was also associated with a significant increase in macrolide-resistant enterococci throughout the GIT. CONCLUSIONS: Dietary acidification is less effective than Ty in modulating the population dynamics of selected culturable populations of enteric bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: The mouse can provide a useful experimental model to examine the effects of new dietary supplements, formulations or regimes on changes in microbial population dynamics, including monitoring for antibiotic resistance.     

Cell-free synthesis of membrane proteins: Tailored cell models out of microsomes

Incorporation of proteins in biomimetic giant unilamellar vesicles (GUVs) is one of the hallmarks towards cell models in which we strive to obtain a better mechanistic understanding of the manifold cellular processes. The reconstruction of transmembrane proteins, like receptors or channels, into GUVs is a special challenge. This procedure is essential to make these proteins accessible to further functional investigation. Here we describe a strategy combining two approaches: cell-free eukaryotic protein expression for protein integration and GUV formation to prepare biomimetic cell models. The cell-free protein expression system in this study is based on insect lysates, which provide endoplasmic reticulum derived vesicles named microsomes. It enables signal-induced translocation and posttranslational modification of de novo synthesized membrane proteins. Combining these microsomes with synthetic lipids within the electroswelling process allowed for the rapid generation of giant proteo-liposomes of up to 50 μm in diameter. We incorporated various fluorescent protein-labeled membrane proteins into GUVs (the prenylated membrane anchor CAAX, the heparin-binding epithelial growth factor like factor Hb-EGF, the endothelin receptor ETB, the chemokine receptor CXCR4) and thus presented insect microsomes as functional modules for proteo-GUV formation. Single-molecule fluorescence microscopy was applied to detect and further characterize the proteins in the GUV membrane. To extend the options in the tailoring cell models toolbox, we synthesized two different membrane proteins sequentially in the same microsome. Additionally, we introduced biotinylated lipids to specifically immobilize proteo-GUVs on streptavidin-coated surfaces. We envision this achievement as an important first step toward systematic protein studies on technical surfaces.     

Influence of environment upon the life history of gudgeon, Gobio gobio (L.): a recent and successful colonizer of the Iberian Peninsula

The growth and the reproductive tactics of gudgeon, Gobio gobio (L.) were assessed in three contrasting streams of the Iberian Peninsula: the Rivers Moros and Ucero, tributaries of the R. Duero, and the R. MatarrańGa, a tributary of the R. Ebro. The R. Duero gudgeon have spread in the last 50 years while their presence in the lower tributaries of the R. Ebro followed the construction of reservoirs in the mid 1960s. The life histories of the Moros and MatarrańnGa gudgeon are very similar; rapid growth in spring, early reproduction (1 -year-old), high reproductive cost, short life-span and trend to semelparity. Both populations are strongly influenced by the severe summer droughts that result in (I) reduction in the condition coefficient, (2) reduction in the growth rate (particularly in the River MatarrańGa), (3) single spawning and (4) no gonad development in the summer. The gudgeon in the Ucero delay maturity, grow slower, are multi-spawners and have a longer life span. All the Duero populations come from the same genetic pool and they are actually connected through the populations living along this river basin, whereas the MatarrańGa population is not genetically linked, we hypothesize that the differences between these populations are environmentally induced.     

Circulating Insulin Concentrations,Smoking, and Alcohol Intake Are Important Independent Predictors of Leptin in Young Healthy Men

Objective : Leptin, an adipocyte-secreted hormone, has been shown to signal the status of energy stores to the brain, regulate energy homeostasis, and mediate the neuroendocrine response to food deprivation. Obesity is associated with increased leptin levels, and several hormones, including insulin and glucocorticoids, have been associated with leptin levels and expression in rodents. Although obesity has been strongly associated with increased leptin in humans, a significant percentage of leptin's variability remains unexplained. The role of endogenous hormones, demographic factors, or certain life-style factors in explaining the residual variability of leptin levels has not yet been clarified. We performed this cross-sectional study to document the relative importance of obesity, lifestyle factor, and endogenous hormones in determining serum leptin levels. Research Methods and Procedures : We measured serum concentrations of insulin, Cortisol, testosterone, growth hormone, and dehydroepiandrosterone sulfate; ascertained anthropometric, demographic, and lifestyle characteristics; and studied these variables in relationship to serum leptin concentrations in a sample of young healthy men. Results : Obesity and alcohol intake were independently and positively associated with circulating leptin concentrations. Additionally, cigarette smoking was negatively and independently associated with leptin concentrations. Finally, serum insulin concentration was an independent hormonal determinant of circulating leptin concentrations, whereas serum testosterone was negatively associated with leptin only by bivariate analysis. Discussion : We conclude that, in addition to obesity, cigarette smoking, alcohol intake, and serum insulin levels are associated with leptin levels in a population of healthy young men.     

Neurite outgrowth from progeny of epidermal growth factor–responsive hippocampal stem cells is significantly less robust than from fetal hippocampal cells following grafting onto organotypic hippocampal slice cultures: Effect of brain‐derived neurotrophic factor

Epidermal growth factor (EGF)–responsive stem cells from both developing and adult central nervous system (CNS) can be expanded and induced to differentiate into neurons and glia in vitro. Because of their self‐renewal and multipotent properties, these cells can potentially provide an unlimited tissue source for neural grafting in neurodegenerative disorders. However, the capability of neurons derived from these stem cells to project axons to distant targets following grafting, thereby enabling the restoration of damaged CNS circuitry, remains unknown. We hypothesize that grafted EGF‐responsive stem cells and their progeny are not competent to project axons into distant target sites unless exposed to specific neurotrophic factors. We compared neurite outgrowth between gestation day 14 primary mouse hippocampal cells and EGF‐generated secondary neurospheres of postnatal mouse hippocampal stem cells, following grafting onto the CA3 region of organotypic hippocampal slice cultures prepared from postnatal rats. Neurite outgrowth from grafted cells was visualized using immunohistochemical staining for the mouse specific antigen M6. Fetal hippocampal cells showed extensive and specific neurite outgrowth into many regions of the slice, including the CA1 region and distant subiculum, by 7 days after grafting. In contrast, neurite outgrowth from neurosphere cells was nonspecific and restricted to the immediate surrounding region after either 7 or even 15 days following grafting. Application of brain‐derived neurotrophic factor (BDNF) (5 ng in 0.5 μL) to slices on day 1 after grafting significantly enhanced neurite outgrowth from neurosphere cells, but overall neurite outgrowth from neurosphere cells remained decreased compared to that from fetal hippocampal cells. These results underscore that EGF‐responsive stem cell‐derived neurons possess limited intrinsic capability for long‐distance neurite outgrowth compared to fetal neurons. However, neurite outgrowth from EGF‐responsive stem cell–derived neurons can be enhanced by treating with specific neurotrophic factors such as BDNF. © 1999 John Wiley & Sons, Inc. J Neurobiol 38: 391–413, 1999     

Estimation of Tissue Construction Cost from Heat of Combustion and Organic Nitrogen Content

Abstract. We present a method for estimating the construction costs of plant tissues from measurements of heat of combustion, ash content, and organic nitrogen content. The method predicts glucose equivalents, the amount of glucose required to provide carbon skeletons and reductant to synthesize a quantity of organic product. Glucose equivalents have previously been calculated from the elemental composition of tissue. We define construction cost as the amount of glucose required to provide carbon skeletons, reductant and ATP for synthesizing the organic compounds in a tissue via standard biochemical pathways. The fraction of the total construction cost of a compound or tissue (excluding costs of transporting compounds) that is reflected in its glucose equivalents is the biosynthetic efficiency ( E _B). This quantity varies between 0.84 and 0.95 for tissues with a wide range of compositions. Using the new method, total construction cost can be estimated to ± 6% of the value obtained from biochemical pathway analysis.
Construction costs of leaves of three chaparral species were estimated using the proposed method and compared to previously published values, derived using different methods. Agreement among methods was generally good. Differences were probably due to a combination of inaccuracy in the estimated biosynthetic efficiency and technical difficulties with biochemical analysis, one of the older methods of determining construction cost.     

Modeling squid axon Na+ channel by a nucleation and growth kinetic mechanism

A kinetic model accounting for all salient features of the Na⁺ channel of the squid giant axon is provided. The model furnishes explanations for the Cole-Moore-like effect, the rising phase of the ON gating current and the slow ‘intermediate component’ of its decaying phase, as well as the gating charge immobilization. Experimental ON ionic currents are semi-quantitatively simulated by the use of only three free parameters, upon assuming that the Na⁺ channel opening proceeds along with the stepwise aggregation of its four domains, while they are moving their gating charge outward under depolarizing conditions. The inactivation phase of the ON ionic current is interpreted by a progressive electrostatic attraction between the positively charged ‘hinged lid’ containing the hydrophobic IFM triad and its receptor inside the channel pore, as the stepwise outward movement of the S4 segments of the Na⁺ channel progressively increases the negative charge attracting the triad to its receptor. The Na⁺ channel closing is assumed to proceed by repolarization-induced disaggregation of its domains, accompanied by inward movement of their gating charge. The phenomenon of ‘gating charge immobilization’ can be explained by assuming that gradual structural changes of the receptor over the time course of depolarization strengthen the interaction between the IFM triad and its receptor, causing a slow release of the gating charge during the subsequent repolarization.     

Synthesis and biological activity of FGLamide allatostatin analogs with Phe3 residue modifications

A FGLamide allatostatin neuropeptide mimic ( H17 ) is a potential insect growth regulator which inhibits the production of juvenile hormone by the corpora allata. To find more evidence to reveal the structure–activity relationships of the Phe³ residue in the C‐terminal conserved pentapeptide and search for novel analogs with high activity, a series of Phe³ residue‐modified analogs were designed and synthesized using H17 as the lead compound. Bioassay using juvenile hormone (JH) production by corpora allata of the cockroach Diploptera punctata indicated that analogs 4 , 11 , and 13 showed strong ability to inhibit JH production in vitro, with IC₅₀ of 38.5, 22.5, and 26 nM, respectively. As well, the activity of analog 2 (IC₅₀: 89.5 nM) proved roughly equivalent to that of H17 . Based on the primary structure–activity relationships of Phe³ residue, we suggest that for analogs containing six‐membered aromatic rings, removing the methylene group of Phe³ or an o‐halogen or p‐halogen‐substituted benzene ring could increase the ability to inhibit biosynthesis of JH. This study will be useful for the design of new allatostatin analogs for insect management. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.     

Cortical bone growth and dietary stress among subadults from Nubia's Batn el Hajar

Cortical bone growth is analyzed for 174 children from a Medieval Christian population at Kulubnarti in the Batn el Hajar of Sudanese Nubia (550–1450 AD ). Using the tibia as a representative long bone, total subperiosteal area, cortical area, medullary area, and percent cortical area at midshaft were calculated. While growth in total and cortical areas, as well as in length, appear to be fairly well maintained, percent cortical area reveals unusual growth patterns which reflect excessive endosteal resorption. Compared to the relative reduction in bone mass which has been observed in malnourished living children, as well as with previously reported evidence for stress in the Kulubnarti population, the present data support an interpretation of nutritionally related stress and of no major diachronic dietary change.