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Dec 30, 2013 - Streptomyces aegyptia NEAE 102 as Eco-Friendly Nanofactory. Noura El-Ahmady El-Naggar1*, ... The biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102 was quite ...... Spirulina platensis. Nanomater.
J. Microbiol. Biotechnol. (2014), 24(4), 453–464 http://dx.doi.org/10.4014/jmb.1310.10095
Fabrication of Biogenic Antimicrobial Silver Nanoparticles by Streptomyces aegyptia NEAE 102 as Eco-Friendly Nanofactory Noura El-Ahmady El-Naggar1*, Nayera A. M. Abdelwahed2, and Osama M. M. Darwesh3 1
Department of Bioprocess Development, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria 21934, Egypt 2 Chemistry of Natural and Microbial Products Department, National Research Center, 12311 Dokki, Cairo, Egypt 3 Agricultural Microbiology Department, Agricultural and Biology Research Division, National Research Center, 12311 Dokki, Cairo, Egypt Received: October 28, 2013 Revised: December 22, 2013 Accepted: December 24, 2013
First published online December 30, 2013 *Corresponding author Phone: +(002)01003738444; Fax: +(002)03-4593423; E-mail: [email protected] com
The current research was focused on the extracellular biosynthesis of bactericidal silver nanoparticles (AgNPs) using cell-free supernatant of a local isolate previously identified as a novel Streptomyces aegyptia NEAE 102. The biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102 was quite fast and required far less time than previously published strains. The produced particles showed a single surface plasmon resonance peak at 400 nm by UV–Vis spectroscopy, which confirmed the presence of AgNPs. Response surface methodology was chosen to evaluate the effects of four process variables (AgNO3 concentration, incubation period, pH levels, and inoculum size) on the biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102. Statistical analysis of the results showed that the linear and quadratic effects of incubation period, initial pH, and inoculum size had a significant effect (p < 0.05) on the biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102. The maximum silver nanoparticles biosynthesis (2.5 OD, at 400 nm ) was achieved in runs number 5 and 14 under the conditions of 1 mM AgNO3 (1-1.5% (v/v)), incubation period (72-96 h), initial pH (9-10), and inoculum size (2-4% (v/v)). An overall 4-fold increase in AgNPs biosynthesis was obtained as compared with that of unoptimized conditions. The biosynthesized silver nanoparticles were characterized using UV-VIS spectrophotometer and Fourier transform infrared spectroscopy analysis, in addition to antimicrobial properties. The biosynthesized AgNPs significantly inhibited the growth of medically important pathogenic gram-positive (Staphylococcus aureus) and gram-negative bacteria (Pseudomonas aeruginosa) and yeast (Candida albicans).
pISSN 1017-7825, eISSN 1738-8872 Copyright © 2014 by The Korean Society for Microbiology and Biotechnology
Keywords: Silver nanoparticles, biosynthesis, antimicrobial activity, Streptomyces aegyptia NEAE 102, response surface methodology, energy dispersive X-ray spectroscopy.
Introduction Nanomaterials have a long list of applicability in improving human life and its environment. An important area of research in nanotechnology is the biosynthesis of nanoparticles such as nanosilver. The extremely small size and large surface area relative to their volume makes them useful for applications in various fields, such as antibacterials [34], therapeutics [40], high-sensitivity biomolecular detection and diagnostics, silver-nanocoated medical devices [37],
optical receptors [26], nonlinear optics, spectrally selective coating for solar energy absorption, biolabeling, intercalation materials for electrical batteries, catalysis in chemical reactions [9], cosmetics, microelectronics, and conductive inks and adhesives [2]. Silver nanoparticles (AgNPs) are currently commonly synthesized by chemical reduction [21], thermal treatment [35], irradiation [30] and laser ablation [38], which are lowyield, energy-intensive, difficult to scale up, often producing high levels of hazardous wastes, and may require the use
April 2014 ⎪ Vol. 24 ⎪ No. 4
El-Naggar et al.
of organic solvents and toxic reducing agents like sodium borohydride and N,N-dimethylformamide. Thus, these techniques yield extremely expensive materials. In addition, the produced nanoparticles exhibit undesirable aggregation with time. Hence, a green synthesis method has been developed to obtain biocompatible, cost-effective, clean, nontoxic, easily scaled up for large-scale synthesis, and ecofriendly size-controlled nanoparticles. Microbial properties of bioaccumulation, biosorption, biodetoxification, and biomineralization have been regarded as opportunities to use them as nanofactories for mining nanomaterials [18]. One of the major issues to be considered during synthesis of nanoparticles is their size, depending on which the applications may vary [7]. Biosynthetic methods can be categorized into intracellular and extracellular synthesis according to the place where nanoparticles are formed [36]. For extracellular synthesis, it is reported that a reductase enzyme is released into solution, which reduces a salt to its metallic solid nanoparticles through the catalytic effect [1]. Extracellular secretion of enzymes offers the advantage of obtaining large quantities in a relatively pure state, free from other cellular proteins associated with the organism, and can be easily processed by filtering of the cells and isolating the enzyme for nanoparticles synthesis from cellfree filtrate. Various microorganisms (bacteria, yeast, fungi) are known to synthesize silver nanoparticles. The produced nanoparticles have different sizes and shapes. The shape of AgNPs, predominantly spherical, is common to microbial-mediated synthesis [31]. Nanoparticles resulting from some microbial processes are composite materials and consist of an inorganic component and a special organic matrix (proteins, lipids, or polysaccharides) and they have unique chemical and physical properties different from the properties of conventionally produced nanoparticles and of other microorganisms even when they are incubated in the same medium under the same conditions [19]. Traditionally, fermentation processes have been optimized by changing one independent variable or factor at a time while keeping the others at some fixed values. The disadvantages of such a classical method are that it is time consuming, laborious, and expensive; in addition, it ignores the combined interactions among different variables employed [6]. Consequently, statistical methods are increasingly preferred for fermentation optimization because they reduce the total number of experiments needed and provide a better understanding of the interactions among factors on the outcome of the fermentation [24]. Response surface methodology (RSM) is a statistical technique based on the fundamental principles of statistics, randomization, replication,
J. Microbiol. Biotechnol.
and duplication, which simplifies the optimization by studying the mutual interactions among the variables over a range of values in a statistically valid manner. It is an efficient statistical technique for optimization of multiple variables in order to predict the best performance conditions with a minimum number of experiments and explain the individual and interactive effects of test variables on the response [20]. In this study, for the first time, a novel actinomycete strain, Streptomyces aegyptia NEAE 102, is utilized for the synthesis of silver nanoparticles. A statistical approach (central composite design) has been employed in the present study to optimize the extracellular biosynthesis of silver nanoparticles by the novel Streptomyces aegyptia NEAE 102. The biosynthesized nanoparticles were characterized for their surface properties using FTIR, in addition to antimicrobial properties using gram-positive, and gram-negative bacterial strains and yeast.
Materials and Methods Microorganisms and Cultural Conditions The Streptomyces sp. used in this study is a local isolate previously identified as the novel Streptomyces aegyptia NEAE 102, and its sequencing product was deposited in the GenBank database under Accession No. HQ677021 [5] and was kindly provided by Dr. Noura El-Ahmady El-Naggar (Department of Bioprocess Development, Genetic Engineering and Biotechnology Research Institute, City for Scientific Research and Technologial Applications, Alexandria, Egypt). This isolate was maintained on slopes containing starch-nitrate agar medium of the following composition (g/l): Starch 20; KNO3 2; K2HPO4 1; MgSO4·7H2O 0.5; NaCl 0.5; CaCO3 3; FeSO4·7H2O 0.01; agar 20; and distilled water up to 1 L. Slopes were incubated for a period of 7 days at 30°C. The isolate was stored as spore suspensions in 20% (v/v) glycerol at –20°C for subsequent investigation. Inoculum Preparation Erlenmeyer flasks (250 ml) containing 50 ml of medium consisting of (g/l): Soluble starch 20; NaNO3 2; K2HPO4 1; MgSO4·7H2O 0.5; and distilled water up to 1 L were inoculated with three disks of 9 mm diameter taken from 7-day-old stock culture grown on starch nitrate agar medium. The flasks were incubated for 48 h in a rotatory incubator shaker at 30ºC and 200 rpm and were used as inoculum for subsequent experiments. Extracellular Synthesis of AgNPs In order to screen an efficient strain for the synthesis of AgNPs, 7 Streptomyces strains (Streptomyces aegyptia NEAE 102, Streptomyces viridochromogenes, Streptomyces sp. NEAE D, Streptomyces albogriseolus, Streptomyces mutabilis NEAE F, Streptomyces sp. NEAE 126, and
Fabrication of Biogenic Silver Nanoparticles
Streptomyces sp. NEAE 25) were freshly inoculated in an Erlenmeyer flask containing 50 ml of the production medium consisting of (g/l): Soluble starch 20; NaNO3 2; K2HPO4 1; MgSO4·7H2O 0.5; and distilled water up to 1 L. The inoculated flasks were incubated on a rotatory incubator shaker at 30°C and 200 rpm for 72 h. After the incubation period, the cell-free supernatant was obtained by centrifugation at 5,000 rpm for 30 min. For the biosynthesis of AgNPs to occur, 1% (v/v) of 1 mM AgNO3 was added to the cell-free supernatant and incubated on an orbital shaker (dark condition) for 24 h at 30°C. The cell-free supernatant without addition of AgNO3 was maintained as a control. Rapid biosynthesis of AgNPs was achieved by addition of silver nitrate solution (1 mM AgNO3) to the cell-free supernatant of Streptomyces aegyptia NEAE 102, where immediately it turned to a brown color solution. Consequently, the bioreduction reaction was monitored by visual color change and UV– visible absorbance of the reaction mixture in the 300–600 nm range. Experimental Design In this study, the effect of four process parameters, namely AgNO3 concentration (X1), incubation period (X2), pH level (X3), and inoculum size (X4), on biosynthesis of silver nanoparticles was studied and optimized using central composite design (CCD). Each factor in the design was studied at five different levels (-2, -1, 0, 1, 2). The coded and actual values of the variables at various levels are given in Table 1. The central values (zero level) chosen for experimental design were 1 mM AgNO3 (1% (v/v)), incubation period 72 h, pH level 8, and inoculum size 4% (v/v). The experimental design used for the study consisted of 30 trials (Table 2). Experiments were conducted in 250 ml Erlenmeyer flasks containing 50 ml of medium prepared according to the design. The flasks were kept in an incubator shaker maintained at 30°C and 200 rpm. All the experiments were done in duplicate and the average of absorbance at 400 nm, which indicates the biosynthesis of silver nanoparticles obtained, was taken as the dependent variable or response (Y). The experimental results of CCD were fitted via the response surface regression procedure, using the following second-order polynomial equation: 2
Y = β0 + ∑ βi Xi + ∑ βii Xi + ∑ βij Xi Xj i
where Y is the predicted response, β0 is the regression coefficients, βi is the linear coefficient, βii is the quadratic coefficients, βij is the interaction coefficients) and Xi is the coded levels of independent variables. However, in this study, the independent variables were coded as X1, X2, X3, and X4. Thus, the second-order polynomial equation can be presented as follows: Y = β0 + β1 x1 + β2 x2 + β3 x3 + β4 x4 + β12 x1 x2 + β13 x1 x3 + β14 x1 x4 2
+ β23 x2x3 + β24 x2x4 + β11 x1 + β22x2 + β33 x3 + β44 x4
Table 1. Process variables used in the central composite design (K = 4) with actual factor levels corresponding to coded factor levels. Coded and actual levels
Variable code
AgNO3, 1 mM (%, v/v)
Incubation period (h)
Initial pH level
Inoculum size (%, v/v)
The graphical representation of the model equation results in response surface plots that represent the individual and interactive effects of test variables on the response. Statistical Analysis The experimental data obtained were subjected to multiple linear regressions using Microsoft Excel 2007 to evaluate the analysis of variance (ANOVA). The quality of fit of the regression model was expressed via the correlation coefficient (R), the coefficient of determination (R2), and the adjusted R2, and its statistical significance was determined by an F-test. The optimal value of activity was estimated using the Solver function of Microsoft Excel tools. The statistical software package STATISTICA software (Ver. 8.0; StatSoft Inc., Tulsa, USA) was used to plot the three-dimensional surface plots, in order to illustrate the relationship between the responses and the experimental levels of each of the variables utilized in this study. Characterization of Silver Nanoparticles UV–visible spectral analysis. The biosynthesized silver nanoparticles using the cell-free supernatant were monitored by changes in color. AgNPs were characterized by a UV-vis spectrophotometer (Jenway UV/Visible- 2605 spectrophotometer, England) scanning in the range of 300–600 nm at regular intervals. Cell-free supernatant without addition of silver nitrate was used as a control throughout the experiment. Fourier-transform infrared spectroscopy analysis. The interaction between protein and AgNPs was analyzed by Fourier transforminfrared spectroscopy (FTIR). The synthesized AgNPs sample was freeze dried and diluted with potassium bromide (in the ratio of 1:100) to make a pellet. The FTIR spectrum of samples was recorded on a FTIR instrument (FTIR-8400S). The measurement was carried out in the range of 500-4,000 cm-1 at a resolution of 1 cm-1. Antimicrobial activity of AgNPs by agar diffusion method. Antimicrobial activity was tested for the biosynthesized AgNPs and cell-free supernatant against bacterial pathogens (obtained from Northern Regional Research Laboratory Peoria, Illinois 61604 USA) of gram-positive (Staphylococcus aureus NRRL B-313) and gram-negative bacteria (Pseudomonas aeruginosa NRRL B-23) and yeast (Candida albicans NRRL Y- 477) using the well-diffusion method on Luria-Bertani (LB) agar plates. A 100 µl bacterial
Table 2. Full factorial central composite design matrix of four process variables and mean response for biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102. Absorbance at 400 nm
suspension of each bacterial test organism was used to prepare bacterial lawns. Agar wells of 6 mm diameter were prepared with the help of a sterilized stainless-steel cork borer. The wells were loaded with 50 µl of AgNPs solution and 50 µl of cell-free supernatant without AgNO3 as a control. The plates were incubated at 30oC for 24 h and then were examined for the presence of inhibition zones. The diameter of such zones of inhibition was measured and the mean value for each organism was recorded and expressed in millimeter unit.
Results and Discussion Extracellular Biosynthesis of Silver Nanoparticles The biosynthesis of silver nanoparticles was performed
by using 100 ml of cell-free supernatant of Streptomyces aegyptia NEAE 102 treated with 1% (v/v) of 1 mM aqueous AgNO3. The reaction mixture was kept in dark to avoid a photolytic reaction. Immediately, the pale yellow colored reaction mixture turned into a brown color within one minute indicating the formation of AgNPs (Fig. 1A). The biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102 was quite fast as in our previous work [4] and required far less time than previously published strains. The time required for completion of nanoparticle production varied from 24 to 120 h; this lengthy reaction is the one major drawback of biological synthesis and must be corrected if it is to compete with chemical procedures for
symmetry of the nanoparticle decreases [33]. Past studies suggested that DNA, sulfur-containing proteins [17], and NADH and its dependent enzymes are possibly involved in the biosynthesis of AgNPs by the bioreduction of silver ion to metallic silver [27]. Chun et al. [3] reported that NADHdependent enzymes play a key role in these biosynthetic and biotransformation reactions as an electron carrier. The UV–vis spectrum (Fig. 1B) clearly indicates the formation of AgNPs in the solution and the band corresponding to the surface plasmon resonance occurs at 400 nm. Ahmad et al. [1] showed that the band corresponding to the surface plasmon resonance occurs at 415 nm, and the exact position of absorbance depends on a number of factors such as the dielectric constant of the medium, size of the particle, etc.
Fig. 1. Analyses of AgNPs produced by Streptomyces aegyptia NEAE 102. (A) Visible observation of AgNPs biosynthesis by Streptomyces aegyptia NEAE 102: (1) Cell-free supernatant. (2) After exposure to AgNO3 solution (1 mM). (B) UV–Vis absorption spectrum of silver nanoparticles synthesized by cell-free supernatant of Streptomyces aegyptia NEAE 102. The absorption spectrum of silver nanoparticles exhibited a strong broad peak at 400 nm, and observation of such a band is assigned to the surface plasmon resonance of the particles.
nanoparticle synthesis [29, 39]. UV-Visible Spectral Analysis In order to determine the formation of extracellular AgNPs, the reaction mixture was monitored by UV–Vis absorption spectrum scanning in the range of 300–600 nm. The specific surface plasmon resonance (SPR) spectra of silver nanoparticles produced by the Streptomyces aegyptia NEAE 102 revealed a strong absorption peak at 400 nm, indicating the presence of AgNPs (Fig. 1B). The position and shape of the plasmon absorption of silver nanoclusters strongly depend on particle size, stabilizing molecules, or surface adsorbed particles, as well as the dielectric constant of the medium [10]. According to Mie's theory [16], only a single SPR band is expected in the absorption spectra of spherical nanoparticles, whereas anisotropic particles could give rise to two or more SPR bands depending on the shape of the particles. The number of SPR peaks increases as the
Optimization of Silver Nanoparticles Biosynthesis Using Response Surface Methodology In order to evaluate the relationship between response and independent variables and to determine the maximum silver nanoparticles biosynthesis corresponding to the optimal levels of AgNO3 concentration (X1), incubation period (X2), initial pH (X3), and inoculum size (X4), a secondorder polynomial model was proposed to calculate the optimal levels of these variables. The results of experiments are represented in Table 2. The results showed considerable variation in the silver nanoparticles biosynthesis. Treatment runs 5, 14, and 26 showed a high silver nanoparticles biosynthesis (≥ 2.3). The maximum silver nanoparticles biosynthesis (2.5) was achieved in runs 5 and 14 under the conditions of 1 mM AgNO3 (1-1.5% (v/v)), incubation period (72-96 h), initial pH (9-10), and inoculum size (2-4% (v/v)), whereas the minimum silver nanoparticles biosynthesis (0.011) was observed in run number 3. By applying multiple regression analysis on experimental data of central composite design, a second-order polynomial model explains the role of each variable and their secondorder interactions on silver nanoparticles biosynthesis. The determination coefficient (R2) values provide a measure of how much variability in the observed response values can be explained by the experimental factors and their interactions. When R2 is closer to 1, correlation is better between the experimental values and the values predicted by the second order polynomial model. The high R2 value implies high degree of correlation between the observed and predicted values [20]. In this case, the value of the determination coefficient (R2 = 0.98138) indicates that 98.138% of the variability in the response was attributed to the given independent variables, and only 1.862% of the total variations are not explained by the independent variables.
Table 3. Analysis of variance for optimization of silver nanoparticles biosynthesis using central composite design. Regression
Significance F (P-value)
2.19E-10
df, Degree of freedom; SS, Sum of squares; MS, Mean sum of squares; F, Fisherss function; Significance F, Corresponding level of significance.
In addition, the value of the adjusted determination coefficient (Adj. R2 = 0.96401) was also very high, which indicates a high significance of the model. A higher value of the correlation coefficient (R = 0.99065) signifies an excellent correlation between the independent variables, which indicated a good correlation between the experimental and predicted values. Thus, the analysis of the response trend using the model was considered to be reasonable. ANOVA was employed to test the significance and adequacy of the model. This was carried by Fisher's statistical test for the analysis of variance. The F-value indicates the influence (significance) of each controlled factor on the tested model; the results are presented in Table 3. The ANOVA of the regression model demonstrated that the model is highly significant, as was evident from the Fisher's F-test (56.4763) and a very low probability value (2.19E-10). The significance of each coefficient was determined by t-values and P-values, which are listed in Table 4. The P-values denote the significance of the coefficients and are also important in understanding the
pattern of the mutual interactions between the variables. It can be seen from the degree of significance that the linear and quadratic effects of X2 (incubation period), X3 (initial pH), and X4 (inoculum size) are significant, meaning that they can act as limiting factors and little variation in their value will alter the product production rate. Furthermore, the probability values of the coefficient suggest that among the four variables studied, X2 (incubation period) and X3 (initial pH) shows maximum interaction between the two variables (0.000), indicating that 100% of the model is affected by these variables, followed by interaction between AgNO3 concentration(X1) and initial pH(X3). On the other hand, among the different interactions, the interaction between X1 (AgNO3 concentration), X2 (incubation period); interaction between X1 (AgNO3 concentration), X4 (inoculum size); interaction between X3 (initial pH), X4 (inoculum size); and linear and quadratic effects of X1 did not show significant effect on silver nanoparticles biosynthesis. In order to evaluate the relationship between dependent and independent variables and to determine the maximum
Table 4. Estimated regression coefficients for optimization of silver nanoparticles biosynthesis using central composite design. Variables
Coefficients
Main effect
t-Stat
AgNO3 concentration (X1)
Incubation period (X2)
Initial pH level (X3)
Inoculum size (X4)
X1 X 2
t, Student's test; P, corresponding level of significance. Multiple, R 0.99065; R Square, 0.98138; Adjusted R Square, 0.96401.
silver nanoparticles biosynthesis corresponding to the optimum levels of AgNO3 concentration (X1), incubation period (X2), initial pH (X3), and inoculum size (X4), a second-order polynomial model (Eq. (3)) was proposed to calculate the optimum levels of these variables. By applying the multiple regression analysis on experimental data, the second-order polynomial equation that defines predicted response (Y) in terms of the independent variables (X1, X2, X3, and X4) was obtained: Y(Silver nanoparticles biosynthesis) = 0.101 + 0.0136 X1 + 0.365 X2 + 0.360 X3 – 0.210 X4 + 0.055 X1 X2 – 0.099 X1 X3 + 0.002 X1 X4 + 0.385 X2X3 – 0.089 X2 X4 – 0.037 X3 X4 + 0.0172 X12 + 0.142 X22 + 0.435 X32 + 0.141 X42 (3) where Y is the predicted response, X1 the coded value of AgNO3 concentration, X2 the coded value of incubation period, X3 the coded value of initial pH, and X4 the coded value of inoculum size. A comparison was made between the experimental and theoretical predictions by the model. The predicted and observed responses are presented in Table 2. This result shows that the predictive accuracy of the model is high. The Pareto chart illustrates the order of significance of the variables affecting silver nanoparticles biosynthesis in the central composite design, as shown in Fig. 2. Among the four variables, the quadratic effect of initial pH showed the highest positive significance by 18.502%. Next to the quadratic effect of initial pH, the interaction between incubation period and initial pH showed a positive effect by 16.377%. The interaction between initial pH and inoculum size showed the higher negative effect by 1.568%.
Fig. 2. Pareto chart illustrating the order of significance of the variables affecting the AgNPs biosynthesis by Streptomyces aegyptia NEAE 102 (the red color represents negative effects and the blue color represents positive effects; Ranks (%) values range from 0.090 to 18.502).
The interaction effects and optimal levels of the variables were determined by plotting the response surface curves (shown in Figs. 3A–3F) when one of the variables is fixed at optimum value and the other two are allowed to vary. Fig. 3A shows that there is gradual increasing of the silver nanoparticles biosynthesis with increasing the levels of AgNO3 concentration. The maximum silver nanoparticles yield appeared at high levels of incubation time. Fig. 3B shows that higher levels of pH support high silver nanoparticles yield. On the other hand, lower and middle levels of AgNO3 concentration support relatively high levels of silver nanoparticles yield. Fig. 3C represents the interaction between AgNO3 concentration and inoculum size. It showed that a low level of inoculum size supports high silver nanoparticles yield and further increase in the inoculum size led to a decrease in the silver nanoparticles biosynthesis. On the other hand, the maximum silver nanoparticles biosynthesis was clearly situated close to the central point of AgNO3 concentrations. Fig. 3D represents the three-dimensional plot as a function of pH and incubation time on silver nanoparticles biosynthesis. It was observed that there is gradual increasing of the silver nanoparticles biosynthesis with increasing both pH and incubation time. The maximum silver nanoparticles yield appeared at high levels of both pH and incubation time. Fig. 3E depicts the interaction of incubation time and inoculum size. The plot reveals that the low level of inoculum size and high level of incubation time supported high silver nanoparticles yield. Fig. 3F represents the threedimensional plot as a function of pH and inoculum size on silver nanoparticles biosynthesis. According to the plot, alkaline initial pH caused maximum silver nanoparticles biosynthesis at a low level of inoculum size. The biosynthesis was decreased remarkably as the pH decreased, whereas further increase in the inoculum size resulted in a gradual decrease in silver nanoparticles yield. A low inoculum may require a longer time for microbial multiplication and substrate utilization to produce the desired product. On the other hand, a high inoculum would ensure rapid proliferation of the microbial biomass. Thus, a balance between the proliferating biomass and substrate utilization would yield maximum activity as recorded by Ramachandran et al. [23]. As the concentration of inoculum increases, it is followed by an increase in cell mass, and, after a certain period, metabolic waste interferes with the production of metabolites due to degradation of the product occurring. A lower inoculum density may reduce product formation, whereas a higher inoculum may lead to poor product formation, especially the large
Fig. 3. Three-dimensional response surface (A-F) showing the interactive effects of independent variables (AgNO3 1 mM (%, v/v), incubation period (h), pH levels, inoculum size (%, v/v)) on biosynthesis of silver nanoparticles.
accumulation of toxic substances, and can also cause the reduction of dissolved oxygen and nutrient depletion in the culture media [22]. The pH value of the cultivation medium is very important for growth of the microorganism, characteristic of their metabolism, and hence for biosynthesis of metabolites. The hydrogen ion concentration may have a direct effect on the cell or it may indirectly affect it by varying the dissociation degree of the medium components [8]. Verification of the Model Optimal concentrations of the factors, obtained from the optimization experiment, were verified experimentally and compared with the predicted data. The measured silver nanoparticles biosynthesis was 2.4 OD at 400 nm, where the predicted value from the polynomial model was 2.32 OD at 400 nm. The verification revealed a high degree of accuracy of the model of more than 96.6%, indicating the model validation under the tested conditions. The optimal levels of the process variables for silver nanoparticles biosynthesis by Streptomyces aegyptia NEAE 102 were 1 mM AgNO3 (1.5% (v/v)), incubation period (96 h), initial pH (9), and inoculum size (2% (v/v)). Antimicrobial Activity of Synthesized AgNPs The antimicrobial activity of synthesized nanoparticles was assessed using gram-positive (Staphylococcus aureus NRRL B-313) and gram-negative (Pseudomonas aeruginosa NRRL B-23) bacterial strains and yeast (Candida albicans NRRL Y- 477) by the well-diffusion method (Fig. 4). A control (cell-free fermentation broth without AgNO3 addition) was also maintained in each plate. It was noticed that the produced nanoparticles displayed antimicrobial activity against selected test microorganisms. The highest antimicrobial activity was observed against Pseudomonas aeruginosa NRRL B-23 (28 mm), followed by Staphylococcus aureus NRRL B-313 (25 mm) and Candida albicans NRRL Y- 477 (25 mm). The negative control (culture broth without AgNO3) did not show any activity (Fig. 4), which further confirms the antibacterial activity of silver nanoparticles. These findings are in agreement with previous studies that examined the antimicrobial activity of AgNPs against Staphylococcus aureus [28]. Metallic silver in the form of AgNPs has been reported as a potential antimicrobial agent. The antibacterial potency of silver is directly proportional to the concentration of silver ions in the solution [12]. A number of possible mechanisms are proposed for the antibacterial activity of AgNPs. Silver ions have been known to bind with the
Fig. 4. Antibacterial activity of silver nanoparticles produced by Streptomyces aegyptia NEAE 102 against: gram-positive bacteria (Staphylococcus aureus), gram-negative bacteria (Pseudomonas aeruginosa), and yeast (Candida albicans).
negatively charged bacterial cell wall, resulting in the rupture and consequent denaturation of cellular proteins, which leads to cell death [13]. The synthesized AgNPs with smaller size can act drastically on the cell membrane and further interact with DNA and causes inhibition of DNA replication [17], causing depletion of intracellular ATP by rupture of the plasma membrane or by blocking respiration in association with oxygen and sulfhydryl (SH) groups on the cell wall to form R S S R bonds, thereby leading to cell death [11]. Matsumura et al. [15] proposed that silver ions interact with the thiol groups of some of the major enzymes and inactivate them. Silver nanoparticles cause damage by possibly interacting with sulfur and phosphorus-containing compounds such as DNA [32]. Fourier Transform Infrared Analysis FTIR spectroscopy was used to characterize the surface chemistry of silver nanoparticles. The FTIR measurement can also be utilized to study the presence of a protein molecule in the solution. Fig. 5 shows the FTIR spectrum recorded from a pellet of potassium bromide and the silver nanoparticle produced by new isolate Streptomyces aegyptia NEAE 102 after 24 h of reaction. The spectrum showed the presence of six bands. The presence of bands at 1,056 and 1,616 Cm-1 in the FTIR spectrum suggests the capping agent of biosynthesized nanoparticles possesses an aromatic
Fig. 6. EDX spectrum showing a peak between 3 and 4 keV, confirming the presence of silver. Fig. 5. FTIR spectrum recorded from a drop-coated film of silver nanoparticles synthesized by Streptomyces aegyptia NEAE 102.
amine group. In addition, the vibrations located at 1,369 and 2,142 Cm-1 are attributed to the C-O stretching mode. Furthermore, the FTIR spectrum of biosynthesized silver nanoparticles also revealed a peak at 3,382 Cm-1 of the stretching vibration of O-H bonds. This type of FTIR spectrum supports the presence of a protein type of compound on the surface of biosynthesized nanoparticles, confirming that metabolically produced proteins act as capping agents during production and prevent the reduced silver particles agglomeration. In fact, the carbonyl groups from amino acid residues as well as peptides of proteins are known for their strong silver binding property. It has been suggested that the stability of the AgNPs generated using cell-free culture supernatants could be due to the presence of a proteinaceous capping agent that prevents aggregation of the nanoparticles [25]. Energy-Dispersive X-Ray Spectroscopy Energy-dispersive X-ray spectroscopy (EDX) is an analytical technique used for the elemental analysis or chemical characterization of a sample. In the current study, for the confirmation of AgNPs, EDX spectroscopy analysis was performed, which confirmed the presence of elemental silver by the signals (Fig. 6). The optical absorption band peak in the range of 3 to 4 keV is typical for the absorption of metallic silver nanocrystallites [14]. However, there were other EDX peaks for Na, C, O, and Cl, suggesting that they were mixed precipitates from the centrifuged cell-free supernatant.
In conclusion, this study revealed that the novel actinomycete S. aegyptia NEAE 102 synthesized silver nanoparticles extracellularly. The produced silver nanoparticles showed single surface plasmon resonance at 400 nm. The silver nanoparticles formed were characterized by UV–Vis spectroscopy and FTIR. This report presents data for the first time on the ability of the cell-free culture supernatant of S. aegyptia NEAE 102 to convert silver nitrate to AgNPs. The biosynthesized nanoparticles displayed a pronounced antimicrobial activity against gram-negative and grampositive bacterial strains as well as yeast.
References 1. Ahmad A, Mukherjee P, Senapati S, Mandal D, Khan MI, Kumar R, Sastry MJ. 2003. Extracellular biosynthesis of silver nanoparticles using the fungus Fusarium oxysporum. Colloids Surf. B 28: 313-318. 2. Akaighe N, MacCuspie RI, Navarro DA, Aga DS, Banerjee S, Sohn M, Sharma VK. 2011. Humic acid-induced silver nanoparticle formation under environmentally relevant conditions. Environ. Sci. Technol. 45: 3895-3901. 3. Chun YJ, Shimada T, Waterman MR, Guengerich FP. 2006. Understanding electron transport systems of Streptomyces cytochrome P450. Biochem. Soc. Trans. 34: 1183-1185. 4. El-Naggar NE, Abdelwahed NAM. 2014. Application of statistical experimental design for optimization of silver nanoparticles biosynthesis by a nanofactory Streptomyces viridochromogenes. J. Microbiol. 52: 53-63. 5. El-Naggar NE, Sherief AA, Hamza SS. 2001. Streptomyces aegyptia sp. nov., a novel cellulolytic streptomycete isolated from soil in Egypt. Afr. J. Microbiol. Res. 5: 5308-5315. 6. Gao H, Liu M, Liu JT, Dai HQ, Zhou XL, Liu XY, et al. 2009.
Medium optimization for the production of avermectin B1a by Streptomyces avermitilis 14-12A using response surface methodology. Bioresour. Technol. 100: 4012-4016. Gurunathan S, Kalishwaralal K, Vaidyanathan R, Venkataraman D, Pandian SRK, Muniyandi J, et al. 2009. Biosynthesis, purification and characterization of silver nanoparticles using Escherichia coli. Colloids Surf. B 74: 328-335. Jain P, Pundir RK. 2011. Effect of fermentation medium, pH and temperature variation on antibacterial soil fungal metabolite production. J. Agri. Technol. 7: 247-269. Kowshik M, Ashtaputre S, Kharrazi S, Vogel W, Urban J, Kulkarni SK, Paknikar KM. 2003. Extracellular synthesis of silver nanoparticles by a silver-tolerant yeast strain MKY3. Nanotechnology 14: 95-100. Krishnaraj C, Jagan EG, Rajasekar S, Selvakumar P, Kalaichelvan PT, Mohan N. 2010. Synthesis of silver nanoparticles using Acalypha indica leaf extracts and its antibacterial activity against water borne pathogens. Colloids Surf. B 76: 50-56. Kumar VS, Nagaraja BM, Shashikala V, Padmasri AH, Madhavendra SS, Raju BD, Rao KSR. 2004. Highly efficient Ag/C catalyst prepared by electro-chemical deposition method in controlling microorganisms in water. J. Mol. Catal. A Chem. 223: 313-319. Lansdown AB. 2006. Silver in health care: antimicrobial effects and safety in use. Curr. Probl. Dermatol. 33: 17-34. Lin YE, Vidic RD, Stout JE, McCartney CA, Yu VL. 1998. Inactivation of Mycobacterium avium by copper and silver ions. Water Res. 32: 1997-2000. Magudapathy P, Gangopadhyay P, Panigrahi BK, Nair KGM, Dhara S. 2001. Electrical transport studies of Ag nanocrystallites embedded in glass matrix. Physics B 299: 142-146. Matsumura Y, Yoshikata K, Kunisaki S, Tsuchido T. 2003. Mode of bactericidal action of silver zeolite and its comparison with that of silver nitrate. Appl. Environ. Microbiol. 69: 42784281. Mie G. 1908. Beiträge zur Optik trüber Medien, speziell kolloidaler Metallösungen. Ann. Phys. 25: 377-445. Morones JR, Elechiguerra JL, Camacho A, Holt K, Kouri JB, Ramirez JT, Yacaman MJ. 2005. The bactericidal effect of silver nanoparticles. Nanotechnology 16: 2346-2353. Narayanan KB, Sakthivel N. 2010. Biological synthesis of metal nanoparticles by microbes. Adv. Colloid Interface Sci. 156: 1-13. Nelly YT, Kirkesali EI, Pataraya DT, Gurielidze MA, Kalabegishvili TL, Gvarjaladze DN, et al. 2011. Microbial synthesis of silver nanoparticles by Streptomyces glaucus and Spirulina platensis. Nanomater. Appl. Properties 2: 306-310. Panwal JH, Viruthagiri T, Baskar G. 2011. Statistical modeling and optimization of enzymatic milk fat splitting by soybean lecithin using response surface methodology. Int. J. Nutr. Metab. 3: 50-57.
21. Peterson MSM, Bouwman J, Chen A, Deutsch M. 2007. Inorganic metallo-dielectric materials fabricated using two single-step methods based on the Tollen's process. J. Colloid Interface Sci. 306: 41-49. 22. Rahman RNZA, Lee PG, Basri M, Salleh AB. 2005. Physical factors affecting the production of organic solvent-tolerant protease by Pseudomonas aeruginosa strain K. Bioresour. Technol. 96: 429-436. 23. Ramachandran S, Patel AK, Nampoothiri KM, Francis F, Nagy V, Szakacs G, Pandey A. 2004. Coconut oil cake: a potential raw material for the production of α-amylase. Bioresour. Technol. 93: 169-174. 24. Revankar MS, Lele SS. 2006. Increased production of extracellular laccase by the white rot fungus Coriolus versicolor MTCC 138. World J. Microbiol. Biotechnol. 22: 921926. 25. Saifuddin N, Wong CW, Nur Yasumira AA. 2009. Rapid biosynthesis of silver nanoparticles using culture supernatant of bacteria with microwave irradiation. J. Chem. 6: 61-70. 26. Schultz S, Smith DR, Mock JJ, Schultz DA. 2000. Single target molecule detection with non-bleaching multicolor optical immunolabels. Proc. Natl. Acad. Sci. USA 97: 996-1001. 27. Senapati S, Ahmad A, Khan MI, Sastry M, Kumar R. 2005. Extracellular biosynthesis of bimetallic Au-Ag alloy nanoparticles. Small 1: 517-520. 28. Shahverdi AR, Fakhimi A, Shahverdi HR, Minaian S. 2007. Synthesis and effect of silver nanoparticles on the antibacterial activity of different antibiotics against Staphylococcus aureus and Escherichia coli. Nanomed. Nanotech. Biol. Med. 3: 168-171. 29. Shankar SS, Rai A, Ahmad A, Sastry MJ. 2004. Rapid synthesis of Au, Ag and bimetallic Au shell nanoparticles using Neem. J. Colloid Interf. Sci. 275: 496-502. 30. Shao K, Yao J. 2006. Preparation of silver nanoparticles via a non-template method. Mater. Lett. 60: 3826-3829. 31. Shivaji S, Madhu S, Singh S. 2011. Extracellular synthesis of antibacterial silver nanoparticles using psychrophilic bacteria. Process Biochem. 46: 1800-1807. 32. Singh M, Singh S, Prasad S, Gambhir IS. 2008. Nanotechnology in medicine and antibacterial effect of silver nanoparticles. Dig. J. Nanomater. Biostruct. 3: 115-122. 33. Sosa IO, Noguez C, Barrera RG. 2003. Optical properties of metal nanoparticles with arbitrary shapes. J. Phys. Chem. 107: 6269-6275. 34. Souza GIH, Marcato PD, Durán N, Esposito E. 2004. Utilization of Fusarium oxysporum in the biosynthesis of silver nanoparticles and its antibacterial activities. In: IX National Meeting of Environmental Microbiology. Curtiba, PR (Brazil). 35. Sun X, Luo Y. 2005. Preparation and size control of silver nanoparticles by a thermal method. Mater. Lett. 59: 38473850. 36. Tang YX, Subramaniam VP, Lau TH, Lai YK, Gong DG, Kanhere PD, et al. 2011. In situ formation of large-scale Ag/
AgCl nanoparticles on layered titanate honey comb by gas phase reaction for visible light degradation of phenol solution. Appl. Catal. B Environ. 106: 577-585. 37. Tomsic B, Simoncic B, Orel B, Zerjav M, Schroers HJ, Simoncic A, Samardzija Z. 2009. Antimicrobial activity of AgCl embedded in a silica matrix on cotton fabric. Carbohydr. Polym. 75: 618-626. 38. Tsuji T, Iryo KN, Watanabe N, Tsuji M. 2002. Preparation of silver nanoparticles by laser ablation in solution: influence of laser wavelength on particle size. Appl. Surf. Sci. 202: 80-85.
39. Vigneshwaran N, Arati Kathe N, Varadarajan PV, Rajan Nachane P, Balasubramanya RH. 2006. Biomimetics of silver nanoparticles by white rot fungus, Phaenerochaete chrysosporium. Colloids Surf. B Interfaces 53: 55-59. 40. Wang XF, Li SF, Yua HG, Yu JG. 2011. In situ anionexchange synthesis and photocatalytic activity of Ag8W4O16/ AgCl-nanoparticle core–shell nanorods. J. Mol. Catal. A Chem. 334: 52-59.
Report "Fabrication of Biogenic Antimicrobial Silver Nanoparticles by ..."


text:
Bryn Mawr residents discuss future
Tom Mattox
For the past year, a handful of Bryn Mawr residents have sketched out a vision of what the neighborhood might look like in 10 or even 20 years.
"Bryn Mawr is going to change," said Vida Ditter, organizer for the Bryn Mawr Neighborhood Association, noting the neighborhood has experienced development pressures recently from outside the community.
In light of these pressures, BMNA formed a land use committee focused on nine large sites where they believe development will have the most impact on the neighborhood's character and uniqueness. The list includes parcels near I-394 and Bassett Creek.
Along the way, committee members discussed and debated the neighborhood's strengths and weaknesses. Committee member Pat Waddick pointed out that I-394 can be viewed negatively because it splits the neighborhood and creates considerable noise. On the other hand, development along the highway could buffer the noise, he said, "And the views of the city are incredible."
Businessperson Mike Palm, who owns property just south of the highway near Penn Avenue, said he hopes to develop his property to take advantage of those views. He also wants the nearby bluffs to be accessible to the public. In June, he told the committee he was happy to work with them.
Michael Martens, another committee member, said he got involved partly because he wanted to bring a citywide perspective to the group. For example, he said the committee looked at medium-density housing for people at different stages in life. "Creating life-cycle housing is good for the neighborhood and good for the city," he said.
So far, much of the discussion has focused on mixed-use development, which combines housing and commercial projects. Ditter said while the committee's recommendations would not be legally binding, it would "make sure developers hear the voices of the neighborhood."
With the help of intern Peng Xu from the University of Minnesota, the committee has put together over a dozen full-color boards with maps and renderings of potential designs.
The committee will make a presentation to the BMNA board at its next meeting, Aug. 13, 7 p.m. at Bryn Mawr Presbyterian Church, 420 S. Cedar Lake Rd.

redpajama_set_name: RedPajamaCommonCrawl


Winners from National Scrapbook Day!
I have the PP "365 Degrees" theme set and the BG "Basics Kraft" theme set, and know she's going to love them. I've been wanting the Girls Paperie "On Holiday," so Julie, you'll have to let me know how you like it when your goodies come in the mail.
She had her choice of any scrapbooking or card sketches book from SCRAPBOOK GENERATION. Gloria chose "Sketches for Scrapbooking, Volume 5." Good choice … you can't go wrong with these books.
Here are the 4 entries received for the Inspire Us contest and what supplies they used that are also sold by MEMORY WORKS.
A sweet layout with some really cute kids by ERICA. She used Jenni Bowlin BUTTERFLIES.
A gorgeous layout by JULIE (winner of game #3!), using QUEEN BEE papers from Pink Paislee and a great sketch from the MY LITTLE SKETCH BOOK blog.
Thank you so much to everyone who participated in my contests for National Scrapbook Day. Let's do it again next year!
Thanks, Erika. Will be so happy to have that sketch book and will use it lots. Enjoyed your contest.
congrats to all the ladies I had a great time, and thank you Erika you are such a wonderful host.
Congratulations to the winners! Thanks for posting my card!


text:
Connect with the AGTS President
AGTS News
We Want to Hear
from You!
Send your latest news
and photos to alumni@agts.edu.
Rapport Alumni Magazine
Encounter: Journal for Pentecostal Ministry
International Journal of Pentecostal Missiology
Audio/Video Resources and Podcasts
Deborah Fegler
Development & Alumni Coordinator
dfegler@agts.edu
Jennifer S. Hall
jhall@agts.edu
alumni@agts.edu
www.agts.edu/alumni
Invest in Student Scholarships
Assemblies of God Theological Seminary
at Evangel University
January/ February 2016
Keeping Alumni Connected
AGTS at Evangel hosts 8th annual Stanley M. Horton Pentecostal Heritage Lectureship series
Dr. Renea Brathwaite ('05) was the featured speaker of the Stanley M. Horton Pentecostal Heritage Lectureship held February 9-11, 2016. He is the Dean of the Ohio Ministry Network and adjunct professor of Theology at Malone University and Hosanna Bible Training Center.
Speaking on the topic of "A re-examination of the theology of William J. Seymour, A Pentecostal Pioneer," Renea engaged students during class sessions and special chapel services. Read more.
Connect with the President
Dr. Hausfeld will be at the following events:
March 6: Preaching at Highlandville First Assembly of God, Highlandville, Mo.
Dr. Hausfeld wrote an article for the winter edition of the Occasional Bulletin, published by Evangelical Missiology Society, entitled, "What are the Missiological Implications of Affirming or Denying that Christians and Muslims Worship the Same God?"
AGTS Discipleship Network launches new blog to offer fresh perspectives on whole-life discipleship
Funded by a generous gift from the Kern Family Foundation, the AGTS Discipleship Network website was created as an interactive communication tool to keep the conversation about Christian discipleship fresh and invigorating. Offering essays and blogs that share how network participants strive to live out their personal faith and ministry, the site welcomes readers and contributors. Read more.
Don't miss the new International Journal of Pentecostal Missiology, Volume 4
Adult, Graduate, and Seminary Studies Open House
Evangel University will be hosting an open house for adult, graduate and seminary studies Tuesday, March 15, in the Riggs Hall Atrium. Open Houses are held every semester and are intended for prospective adult and graduate students with a desire to complete their undergraduate degree or pursue a master's or doctoral degree in a convenient format. This come-and-go event is a great way to visit facilities, network with faculty and learn more about admissions procedures. Evangel waives the $25 application fee for attendees. Learn more or register.
New Hogan Professor Sobhi Malek to deliver inaugural lecture
AGTS is pleased to announce that Dr. Sobhi Malek has been chosen as the 2016-2017 J. Philip Hogan Professor. Dr. Malek will present three lectures in Seymour Chapel on the general theme of "The Challenge to Understand Islam."
Inaugural lecture: Tuesday, March 8 at 11 a.m.
"The Caliphate: The Islamic State"
Second lecture: Friday, March 11, at 2 p.m.
"Muhammad in the Bible: Alleged Prophecies"
Third lecture: Wednesday, April 6, at 11 a.m.
"The Bible for Muslims"
As an ordained minister with the Assemblies of God, Dr. Malek has ministered for over forty years in over fifty countries. He has served as a visiting professor of missions at several seminaries, and speaks in conferences and seminars on Islam, Muslim evangelism, and missions to Muslims. In addition, Dr. Malek has translated the Bible into Islamic Arabic. The Sharif Bible was the first Bible to be published in Arabic that Muslims understand.
The J. Philip Hogan Chair of World Missions at AGTS is an endowed professorship that honors the missionary leadership of the former executive director of AGWM. As a partnership between AGTS and AGWM, it calls on today's missionaries and scholars to continue in the heritage of thoughtful, incisive and Spirit-led missiology. A leading missiologist is invited annually to fill the chair in order to explore new dimensions in missiology through teaching, research and writing.
AGTS Offers Church Business Administration Training
In association with The Church Network, an interdenominational professional association, AGTS offers training seminars every summer in church administration. Church leaders, paid and unpaid, have an opportunity to enhance their leadership in human resources, finance, communication, strategic planning, tax and legal issues and property and facilities management.
June 14-17 (Module A: HR, Communication, Planning)
June 21-20 (Module B: Finance, Legal, Facilities)
Learn more or register.
Libraries in a bottle: CBC, Evangel, & AGTS books resourse Bible schools across Africa
Recently, Africa's Hope, a ministry of Assemblies of God World Missions, Africa region, received more than 55,000 textual resources from Evangel University (EU) in Springfield, Missouri, as a result of the 2013 consolidation of EU with Central Bible College and Assemblies of God Theological Seminary. This donation served to provide library resources for several Africa AG Bible schools. Read more.
AG scholarships key to helping AGTS chaplaincy candidates reach the field
Missouri Army National Guard Assemblies of God chaplain candidate Peter K. Pyo is scheduled to graduate from Assemblies of God Theological Seminary in May, after which he expects to go on active duty.
Pyo is one of 196 U.S. Missions AG Chaplaincy Ministries candidates to be awarded a total of 610 scholarships to attend the Springfield seminary since 1983. To date, Chaplaincy Ministries has given $1,026,368 to help AGTS students who are in full-time preparation for the chaplaincy. Read more.
Connect to EU Alumni Newsletter---The Quad
Stay up-to-date with what's happening at Evangel University.
Read the January, 2016, edition.
Do you know someone interested in attending AGTS? Use this online form or connect with Christina Ketchem, AGTS Enrollment Representative.
Roger Cotton
March 2: Attending Chi Alpha meeting, Q & A Topic: "Old Testament: God of Wrath," College of the Ozarks, Point Lookout, Mo.
March 13-14: Attending the Central States Regional Meeting of the Society of Biblical Literature in St. Louis, Mo.
March 10-12: Attending the 45th Annual Meeting of the Society for Pentecostal Studies at Life Pacific College, San Dimas, Calif.
Panel participation:
Volume in honor of Ruth and Stan Burgess
Volume on ethics (Essay topic: International Ethics)
Presenting Essay:"Worship and Spirituality in the First Millennium Church of the East Mission in China: Lessons for the Pentecostal Mission Work Today"
Chairing: Mission Interest Group
Johan Mostert
Assumed the position of Director of Family and Community Resources at COMPACT Family Services, a part of the AG Family Services Agency, on February 1, 2016. Dr. Mostert will head up COMPACT's resource and empowerment strategy to better assist and mobilize local churches and AG districts to more adequately redeem and serve vulnerable children and families in their communities. He will also assist in the enhancement of the agency's professional programs. For additional details, visit COMPACT'S website.
Lois Olena
March 10-12: Attending the 45th Annual Meeting of the Society for Pentecostal Studies at Life Pacific College, San Dimas, Calif. (This is her final year of a five-year term as SPS executive director.)
Presentation participation:
Co-edited book, Children of the Calling: Essays in Honor of Stanley M. Burgess and Ruth V. Burgess
Upcoming volume of Global Pentecostal and the Charismatic Studies series
"Building Bridges to Your Jewish Neighbor at Hanukkah." Vital Magazine, December 3, 2015
Ava Oleson
"Women in Pentecostal and Charismatic Ministry: Information and Ongoing Gender-Focused Dialogue on the Faith Contributions of Women"
Delonn and Valerie Rance
March 2-6: Speaking at the Assemblies of God "Together in Missions" Regional Congress, which includes Hispanic Districts in the U.S. and 20 Latin American countries.
Randy Walls
Feb 18-20: Attending the Association of Theological Schools Peer Group on International Education Models, Pittsburgh, Penn.
Craig Keener's ('85/'87) four-volume Acts commentary won the award in biblical studies this year for Christianity Today's 2016 Book Awards. Gary Burge, professor of New Testament at Wheaton College writes,
"Keener is a scholar with gifts that come along once every century, and here we see them employed in full force. Words like encyclopedic, magisterial, and epic come to mind when you examine 4,000 carefully argued pages on every aspect of the Book of Acts. Nothing like this has ever been done--and it's doubtful that anything like it will be done for a long time. Keener has a grasp of the ancient world like few scholars anywhere, but he also has a heart for the church and its mission."
Bill Parks ('08) passed away in January after a long battle with cancer. AGTS professor Jim Hernando writes,
"Bill came to AGTS to pick up a few more tools and sharpen a few others. While in Springfield he made connection with another alum, Everett Ward, director of the LACLS and put together a plan to help the rural pastors of Nicaragua, a Latin American country he deeply loved. Shortly after graduating and taking a pastorate in Crawfordville, Ind., he invited me to teach in Nicaragua. I accepted, went and have been going back yearly ever since. Bill's love for the people of Nicaragua was infectious. He raised tens of thousands of dollars to buy books for the pastors who attended our two-day seminar on basic principles of biblical interpretation."
Greg Stone ('95) is the associate pastor of Jewish Ministries at Gateway Church in Dallas, Texas. Watch his testimony.
Scott Stripling ('96) serves as the director of excavations at Khirbet el-Maqatir in Israel. Khirbet el-Maqatir is believed by many evangelicals to be the site of Ai and Ephraim. Visit the dig website. A new Logos course entitled Archaeology 101, which prominently features Scott and his dig is also available.
Chaplain David Wright ('15) and his call to have a maximum spiritual impact in the military was the subject of the the 2015 fall edition of Evangel University's Vision magazine. Check it out.
AGTS Spring Study Week
Adult, Graduate Studies & Seminary Open House
March 15, 5-7 p.m., Riggs Hall Atrium
Good Friday/AGTS Offices Closed
AGTS Graduation Events
May 5-7
Do you have family or ministry news you want to share? Send your recent news and photos to alumni@agts.edu and we may publish it in the next newsletter, Rapport magazine, or in our "Alumni Spotlight" website feature.


If you are already operating in the world of eCommerce, or just beginning to develop plans for a sales oriented website, you may be surprised by some of the facts and figures associated with eCommerce. For instance, "the online market is growing rapidly, with retail eCommerce sales predicted to top $4 trillion by 2020." That means you have a lot going for you in terms of audience size and potential, but if you have poor eCommerce website design, you are severely reducing your chances.
Though it may initially seem impossible or unreasonable that good prices and a desired product or service won't sell if your eCommerce website design is not the best, the simple truth is that consumers respond to specific types of visual content. With the human brain committing roughly 20% of its resources to sight and vision, it is a good idea to pair with the best web design company to tackle your eCommerce website design or redesign.
As you learn from the experts, "your virtual storefront is the face of your business, and how it looks will affect buyer's behaviors and brand perceptions." It is not merely about being able to buy and sell around the clock. It is not only about making it easy for a customer to find what they want. It is also about making certain that the entire process is quick and easy, that the site is a good "fit" to the mission and focus of the business, and that it establishes a positive brand perception from the moment a visitor arrives.
These are but a few key reasons to work with the best web design company, but another is that they will be well aware of the most significant best practices where eCommerce website design is concerned.
1. Design considerations – Eye catching design that is pleasurable to your specific audience is the best policy here. If it is all about bells and whistles, it is not the best web design company for you.
2. Responsive – It goes without saying that more than half of all consumers shop via a mobile device. If you are unable to enjoy responsive eCommerce website design, look elsewhere.
3. Emphasize product – A nice looking site is great, but it has to channel attention to whatever it is you are selling. The best web design company will do the photography, the site search tools, the "highlights" of best sellers, and everything else needed to ensure sales.
4. Tell the story – Good marketing begins with your story. A good design team should be able to help you with that, as well as support it with their work.
At Arctic Empire, you are working not only with the best web design company for eCommerce, but also encountering a full-service site. Whether it is the SEO to get people to the site, the creative content, the brand positioning and strategy, the development to make sure you are mobile, or any number of web services, you can quickly find what you need.


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Browns West India Quay
Unit A Hertsmere Road, West India Quay, London, E14 4AY 020 7987 9777
10am-11pm 9am-11pm 9am-11pm 9am-11pm 9am-Midnight 9am-Midnight 9am-10pm
A warm welcome awaits you at Browns West India Quay.
Housed in a Grade 1 listed warehouse once used to store imported sugar, Browns West India Quay is steeped in history, occupying a prime spot on the North Dock of Canary Wharf amidst its maritime heritage and modern architecture.
Ideally positioned for relaxation and refreshment after a walk around this fascinating area of London or a visit to one of the capital's myriad attractions, Browns West India Quay offers a vibrant brasserie and bar experience, with exposed original beams and brickwork characterising the comfortable interior. Outside, a generous terrace provides a peaceful waterside spot for al-fresco drinking and dining.
From breakfast to dinner, afternoon tea to à la carte, our menu delivers fresh and seasonal dishes to suit any occasion or party size, whilst our exceptional drinks range, including classic and contemporary cocktails, global wines and artisan spirits, provides reason enough to relax and unwind at our welcoming bar.
Friday 9am-Midnight
Saturday 9am-Midnight
Browns West India Quay, Unit A Hertsmere Road, West India Quay,
Greater London, UK, E14 4AY
Unit A Hertsmere Road, West India Quay, London, Greater London, E14 4AY


Get in the game! At Oakton Community College, you'll have plenty of opportunities to get involved in athletic activities. Whether you'd like to join an intercollegiate sports team, play at the intramural level, or work out on your own, it's easy to get involved.
As a member of the NJCAA and Illinois Skyway Collegiate Conference, Oakton students participate in 12 intercollegiate sports, including baseball, basketball, soccer, tennis, and golf.
From table tennis to basketball, intramurals at Oakton are a great way to get involved and stay active. Intramurals are open to all students and employees of the College, and you can join as an individual or with a team.
Located at the Des Plaines campus, Oakton's Fitness Center features more than 20 workout machines and an experienced staff.
Make sure your workout is working for you. Oakton's personal trainers will meet with you to customize an exercise regimen that fits your individual needs.


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Low-cost flights with dependable airways – we know what's vital to you while you search for your flight booking with us. At we recognise that your flight reserving is the start of what could grow to be probably the greatest holidays you will have ever had, so we always goal to provide you with the perfect low cost flights and cheap tickets to your chosen holiday destination.
The very best rated WordPress theme is now on SALE! Professionals: Among the greatest at ferreting out the bottom fares; exhibits worth for similar flight from multiple OTAs/airline websites; can see a whole month of fares on a calendar for flexible dates; exhibits which airline truly operates codeshares; fare alert emails; finds lowest fares to many locations at once with all over the place" characteristic.
Because the variety of aggregators have grown, it's turn out to be tougher to pinpoint the accountable social gathering when a guest has a problem with a booking. You can all the time access these websites in your phone's internet service however an app is sweet, and a few apps are notably helpful as a result of they make booking simpler than it is on a cell website.
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And for good reason – the rampage of vacationers visiting websites which have develop into famous on the 'Gram and spending more time on their filters than on taking a look at a vacation spot actually grinds my gears. A vacation is one thing you need to ensure you're paying for precisely what you get, and meaning reserving by a web site you trust.
Village Travel makes reserving a vacation straightforward and convenient. The data on this Site is common in nature and isn't supposed as an alternative to competent legal advice. The theme park capital of the world has no shortage of occasions this holiday season – together with one thing for all ages. It is permitted to operate a branch of the operation of a tour operator or travel agency on the premise of the licence, given that the department director also fulfil the stipulations listed in the following sections a-e.
Here's the true shift: For the higher part of two decades, journey brokers claimed they worked for you but their loyalties remained with the suppliers, who paid them beneficiant bonuses that lined their expenses. A user name and password is required to entry the ASTA's member restricted E-Library where most of the resources are located.
A travel agent may match for a travel agency or work freelance 23. Resort Life Maldives is one of the leading travel and tour operators in Maldives; contributing to the country's tourism sector since 2006. Journey agencies charge charges for services offered to shoppers, for time spent researching and for booking journey preparations.
F. current a affirmation of legal responsibility insurance coverage from an insurance coverage company. WikiProject Journey and Tourism (or its Portal ) might be able to help recruit an skilled. With us, typical holidays turn into memorable adventures all imbued with inspiring discovery, native connections and authentic experiences.
A travel web site is a website on the world large web that is devoted to travel The positioning could also be targeted on journey reviews , trip fares, or a mixture of both. Hotel comparisons: Tools that will help you evaluate your options will make it simpler so that you can select lodging which can be best for you. By offering robust tools that tremendously simplify user expertise, buyer favorite provides quick access to one of the widest picks of each traditional motels and tougher-to-find lodging choices.
Find it irresistible or hate it, TripAdvisor is the daddy of lodge overview web sites," says Mark. Users can view their itinerary on a map which is nice to find other locations nearby. Aggregators' enterprise models embrace getting feeds from major OTAs, then displaying to the customers all of the outcomes on one display screen.
Guides: When you may have spare time, use cell journey guides to find interesting locations close to you. I clicked on the worth and was redirected to JetBlue Holidays, which had the price at $1,306. In 2015, Lufthansa Group (including Lufthansa, Austrian Airlines, Brussels Airlines and Swiss) announced including surcharge for flights booked on different websites.
Travelocity's first nonsponsored suggestion was a room with a king mattress at the Hard Rock Hotel Chicago, a decent alternative, for $149 per night time. Travelocity affords some nice choices for vacationers, together with a $50 credit and refund of the difference if you could find a cheaper value on a flight, rental car or cruise inside 24 hours of reserving.
Examine costs on 400+ airlines and travel sites, including Expedia, and more. EDreams will mechanically apply the costs with the corresponding discount for residents. Holiday firms' personal insurance coverage is usually an enormous rip-off and presents more restricted cowl. Skyscanner compares hundreds of thousands of flights to seek out you the cheapest deal, fast.
For extra cabin choices, first select particular cities and journey dates. Search, select and save – the fastest technique to guide your trip. We have already performed that by looking lots of of them for you- scouring premium airways, low-cost carriers and the most important online journey agencies for the very best deals.
Please examine your entry or proceed along with your choice on our route network. Examine fares simply and guide the perfect offers on low cost home and worldwide flights. Nevertheless Momondo's figures are based mostly on the costs quoted in flight searches, and that elements in sales too, so take its 'perfect day to ebook' information with a pinch of salt, and if doubtful book early.
Flight brokers may give huge reductions for those who're reserving a hotel as effectively. Find the most effective offers on last minute flights to destinations worldwide. Most of the flights and flight-inclusive holidays on this web site are financially protected by the ATOL scheme.
There are numerous travel agents and an equal member of travel portals providing you low-cost and economical flight tickets if you will take advise of them as a traveler then they are going to suggest you to take a tickets from them however many of the traveler they do not full fill there commitment however sure there is some traveler who all the time present adequate information to they're traveler. In summary, the lengthy-held perception that it's better to buy spherical-trip tickets at any time when attainable to get the perfect fares, is solely now not true," the examine mentioned. Yes, the planning can actually take some time when you get into it. A few cheaper origin cities would come with ICN, DPS, and NRT (I think – not 100% certain if NRT is cheaper now).
Book your resort and flights together and make the most of our biggest reductions! Please overview our Unaccompanied Minor Policy earlier than you proceed with this booking. Join us on Fb for travel chat, sizzling offers and giveaways. Browse here to find the very best Rental Automobile Offers in some of our prime journey places.
Not solely does Opodo offer aggressive prices and maximum comfort but, prospects can enjoy deciding on flights from amongst 1000's of routes to prime locations worldwide, all at great prices. Nicely, they alter once in a while, but as of the time I'm scripting this (December 2008), the perfect places are Korea, South Africa, Mauritius, Sri Lanka, and Indonesia.
Via our relationships with probably the most revered airways in the world, we are able to offer our clients unique affords, seat gross sales, and great offers unavailable elsewhere. You can also arrange electronic mail alerts to get updates on value adjustments.


text:
Southeast Warren Baseball
Winning Streak: W2
Conference Record: 12-3
05/20/19 Murray Box Score W 15-0 Lacona Ball Field
05/22/19 Twin Cedars Box Score W 12-3 Lacona Ball Field
05/23/19 7:00PM East Union Box Score W 25-0 East Union Middle-High School
05/29/19 7:00PM Earlham Box Score W 5-1 EARLHAM SENIOR HIGH SCHOOL
05/30/19 Pella Christian Box Score W 3-2 Lacona Ball Field
06/04/19 6:00PM Martensdale-St Marys Box Score W 3-2 Lacona Ball Field
06/05/19 Lamoni Box Score W 17-6 George Foreman Park
06/07/19 Bedford Box Score W 11-4 Lacona Ball Field
06/10/19 Ankeny Christian Box Score W 15-1 Cownie Baseball Field, Des Moines
06/12/19 Melcher-Dallas Box Score W 15-5 Lacona Ball Field
06/13/19 Wayne, Corydon Box Score W 6-1 Lacona Ball Field
06/14/19 Mount Ayr Box Score W 9-2 Lacona Ball Field
06/17/19 Moravia Box Score W 5-4 Lacona Ball Field
06/17/19 Moravia Box Score W 13-5 Lacona Ball Field
06/18/19 East Union Box Score W 12-2 Lacona Ball Field
06/20/19 Central Decatur, Leon Game Preview Lacona Ball Field
06/21/19 7:30PM Southwest Valley Game Preview Lacona Ball Field
06/24/19 Seymour Box Score W 13-0 Lacona Ball Field
06/25/19 6:00PM Lynnville-Sully Box Score W 16-1 Lynnville-Sully High School
06/26/19 West Central Valley Box Score W 19-1 Lacona Ball Field
06/27/19 Central Decatur, Leon Box Score W 8-1 Lacona Ball Field
06/29/19 Fairfield Box Score L 8-7 Fairfield Middle School
06/29/19 Mediapolis Box Score W 10-2 Fairfield Middle School
07/01/19 Mormon Trail Box Score W 12-0 Mormon Trail Jr-Sr High School
07/02/19 Wayne, Corydon Box Score W 16-1 Unknown
07/03/19 Southwest Valley Box Score W 14-0 Lacona Ball Field
07/05/19 Martensdale-St Marys Box Score L 4-3 Martensdale-St Marys Jr-Sr High School
07/08/19 Eddyville-Blakesburg-Fremont Box Score L 8-7 Eddyville-Blakesburg- Fremont Jr/Sr High
07/09/19 Pleasantville Box Score L 11-7 Pleasantville High School
07/13/19 7:00PM Class 1A District 15 Box Score W 12-0 Milo Ball Park
07/20/19 7:00PM Class 1A District 15 Game Preview Milo Ball Park
Austin Clendenen 14 Sophomore
Kole Manser 11 Junior
Dominic Wadle 8th
Bryce Hall 2 Junior
Colby Page 15 Senior
Kaleb Bauer 3 Senior
Luke Lane Freshman
Jt Rowe 8 Junior
Seth Chumbley 12 Junior
Brock Manser 5 Freshman
Dylan Ruggles 8th
Tanner Dierking 6 Sophomore
Cade Nelson 1 Freshman
Ben Wickett 8th
Lane Dittmer 7 Senior
Jeffrey Oakley Freshman
Jack Williams 8th
Brock Kyner 13 Senior
Will Prater 8th
Ben Crall 8th
Mason Merfeld 4 Sophomore
James Wendt 16 Sophomore
05/20/19 19 8 15 2 0 0 10 10 6 vs Murray
05/22/19 27 11 12 3 1 0 9 10 3 vs Twin Cedars
05/23/19 26 10 25 3 0 1 24 3 13 @ East Union
05/29/19 24 7 5 1 0 1 5 4 4 @ Earlham
05/30/19 26 4 3 2 0 0 1 0 5 vs Pella Christian
05/31/19 33 10 10 2 0 1 7 5 5 @ Lenox
06/05/19 33 16 17 3 1 0 16 16 8 @ Lamoni
06/07/19 31 17 11 1 0 0 10 3 7 vs Bedford
06/10/19 27 12 15 3 0 2 13 5 8 @ Ankeny Christian
06/11/19 34 15 9 2 1 0 8 5 0 @ Central Decatur, L
06/12/19 18 10 15 2 0 1 11 15 12 vs Melcher-Dallas
06/13/19 25 8 6 2 0 0 6 3 3 vs Wayne, Corydon
06/14/19 27 13 9 0 1 0 9 7 2 vs Mount Ayr
06/17/19 31 11 5 4 1 0 5 4 1 vs Moravia
06/17/19 27 11 13 4 0 0 11 7 9 vs Moravia
06/24/19 23 9 13 0 0 1 10 10 5 vs Seymour
06/25/19 28 17 16 3 1 1 16 7 3 @ Lynnville-Sully
06/26/19 22 12 19 3 0 1 18 3 10 vs West Central Vall
06/27/19 23 7 8 1 0 1 7 4 5 vs Central Decatur,
06/29/19 29 8 7 1 0 0 5 0 2 vs Fairfield
06/29/19 30 10 10 1 0 1 9 1 3 vs Mediapolis
07/01/19 28 13 12 2 0 2 12 8 1 @ Mormon Trail
07/02/19 27 17 16 2 0 3 14 2 7 @ Wayne, Corydon
07/03/19 22 7 14 2 0 0 11 3 2 vs Southwest Valley
07/05/19 24 6 3 1 0 0 3 0 7 @ Martensdale-St Mar
07/08/19 25 4 7 2 0 0 5 1 9 @ Eddyville-Blakesbu
07/09/19 28 10 7 6 0 0 6 1 4 @ Pleasantville
07/13/19 20 7 12 1 0 0 8 8 8 vs Orient-Macksburg


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Napoli vs. Lazio 8/1/20 Serie A Soccer Prediction and Picks
Home SOCCER Picks
Chris Ruffolo July 30, 2020 5:51 pm
Lazio (24-6-7) at Napoli (17-8-12)
Serie A Soccer: Saturday, August 1, 2020 at 2:45 PM (Stadio San Paolo)
The Line: Napoli +200 / Draw +270 / Lazio +120 -- O/U: 3 Click Here for the Latest Odds
Where to Watch: ESPN+, DAZN Stream live sports and exclusive originals on ESPN+. Sign up now!
Napoli plays host to Lazio in Serie A soccer action from the Stadio San Paolo on Saturday afternoon.
Napoli comes into this match cemented in 7th place on 59 points with a record of 17 wins, 8 draws and 12 losses following a 2-0 loss to Inter Milan in their last match. In the loss to Inter Milan, Napoli fell behind in the 11th minute and would hold 56% possession in the second half, but failed to record a shot on target in the second 45 while Inter doubled their advantage in the 74th minute to lock up the result and send Napoli back empty-handed and slamming the door on their potential hopes for Europa League for next season. Arkadiusz Milik leads Napoli in scoring with 11 goals while Dries Mertens has 9 goals with 5 assists and Lorenzo Insigne has 7 goals with 6 assists. Jose Callejon has a team-high 7 assists for Napoli this season.
Lazio on the other hand, sits in 4th on place on 78 points with a record of 24-6-7 after a 3rd straight win in the form of a 2-0 win over Brescia last time out. In the win over Brescia, Joaquin Correa gave Lazio the lead after just 17 minutes of play and the goal held up through the half and well into the second half before Ciro Immobile scored again to double Lazio's lead and send them to a 3rd straight win, cruising to the finish line. Immobile leads Serie A in scoring with 35 goals while Luis Alberto has a team-high 15 assists this season.
I considered the plus money with Napoli, but the reality is that this game is for pride and nothing else for Napoli as they're locked into 7th. Lazio can finish as high as 2nd with a win and a lot of help and it doesn't hurt that Lazio's in some pretty good form right now having won three straight coming into this match. I'll take a shot with Lazio to pick up the win on the money line and make things interesting at the top on the final matchday.
Chris's Pick Lazio +120
Brescia vs. Sampdoria 8/1/20 Serie A Soccer Prediction and Picks
San Jose Earthquakes vs. Minnesota United FC - 8/1/20 MLS Soccer Picks and Prediction
AUTHOR: Chris Ruffolo
Chris has a proven system that combines multiple analytical and situational statistics and trends to give out thousands of winning picks over the last three years with winning records across all of the top leagues,
Follow Chris Ruffolo on Twitter @ruffthepickdawg


text:
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assert.sameValue(obj.accProperty, 11, 'obj.accProperty');
assert.sameValue(public, 1, 'public');

redpajama_set_name: RedPajamaGithub


What is project management? Simplest put, the art and science of taking an idea to reality by planning and managing who does what, when, and how. That's the basic definition – but the devil is always in the details, as they say. Project management is about projects, and projects aren't just anything.
Say you're an industrial lighting company. You've won the bid to install all the lighting in a mall. You have a goal – installing the lighting in the mall. This is a unique goal. You have a timeframe – the time between now and the deadline when the work needs to be completed. This is not an ongoing operation.
On the other hand, say your industrial lighting company is given a maintenance contract for the same mall. You are tasked with replacing all the lighting fixtures that burn out or break. This is an ongoing operation – there is no end. This is NOT a project. There is a goal, but there is no timeframe.
OK, you say. A project is an endeavor with a goal and a timeframe. Now that you have a project, you need to manage it to a successful conclusion. In our example, this is a mall with lighting installed, functional and ready to go, by the date of the deadline. How will you accomplish this?
Project management has a few defining factors of its own.
Project management is a top down process – it starts with the goal, and gets broken down to tasks. It's actively managed, against a rubric. Chaos is the enemy of the project.
The first step to managing a project is creating the project plan. A project plan is almost always a timeline – this timeline is usually a Gantt chart, sometimes created as an Excel template or with project management software. This is the "top-down" process – the plan is where the goal is translated into tasks.
This plan, once approved, becomes a tool for managing for the project as it unfolds. Tasks are assigned to team members, progress is reported, milestones are met (or delays are accounted for). Project managers monitor the overall health and progress of the project, delegating workflow appropriately and managing crises as they arise. This is the "execution" and "monitoring" aspects of a project.
What Goes Into Managing a Project?
Quite a lot goes into managing a project. The description sounds idyllic. It might even give you the picture of a paint by numbers approach. It's easy to pretend a project manager just whips up some Gantt charts and overlooks the project, but this isn't true. Reality is anything but. Projects have a lot of moving parts. They're dynamic, and fluid. Trying to impose order, or even just stick to a plan, is difficult. There are a lot of things on a project manager's plate!
The scope of a project is its measurables. This is usually defined in the planning stage. Projects can often begin to act like runaway trains, in that the goal begins to change over time. Without active, ongoing management of the project's scope during the execution phase, that project to install lighting in a new mall can quickly become a contract to wire the entire city block. Who keeps a project nailed to the scope set out at the beginning? The project manager.
Time is of the essence, they say. Managing time in terms of project management is a multi-layered endeavor. You need to manage the project's time, making sure the goal is accomplished before the deadline. Then there's managing the time of the people working on the project, making sure they have enough time to finish their tasks while not becoming wasted resources. Finally, you need to manage the time necessary for tasks to be completed. Who manages the different timelines? The project manager.
Many projects require multiple teams, with multiple members, working on different aspects of the project. Someone has to make sure everyone is on the same page! In our example, one team is on site at the mall installing fixtures, and another is dealing with vendors at multiple external sites. A third is coordinating shipping and warehousing. Yet another is interfacing with the customer. Who is coordinating everything? The project manager is responsible for integrating all the different parts of the project, so they function together.
Bob in procurement sent a message by email saying fixture #442 is on temporary backlog from the supplier. Thing is, Molly in installation works primarily through Slack. She didn't see the message in time. Oh, and Kevin at the warehouse sent his progress report through the project management software system. How do multiple people speak to each other in a way that messages are received and actionable in the right time window? Someone needs to communicate with stakeholders, communicate with the team, and with management. The project manager sets communication channels, and acts as the central nervous system for the project.
Things cost money. Projects are assigned budgets, and someone needs to make sure money is spent properly. Supplies, software, miscellaneous expenses…they all add up! There's labor costs to keep track of, too. Your remote office bought a pen? Someone is paying for that, right? The customer does not want his project to run over budget. It's that simple! Who is tracking the budget, expenses, and labor costs? The project manager.
Sometimes surprises are nice. A surprise birthday party, for example. A bonus in your paycheck. But a lot of surprises are not nice at all. When clients and stakeholders find out that their project is at risk, they aren't happy. With any undertaking, there are risks. Delays, vendor issues, or the unforeseeable – who knows? There's already risk baked into any project. Identifying what the risks are, and taking the preventative steps, is imperative. Creating a robust project plan capable of dealing with crises and sudden risks is crucial. The project manager is the chief risk officer of any project.
Project management isn't as simple as just deciding who does what, when, and how. Project management starts with defining goals and timeframes. It incorporates a top down process. Project management starts with planning, moves to execution, and is monitored throughout. It manages scope, communication, budget, risk, time, and integration.
The best way to think of project management is an airline route map.
There are many inputs, but they all either end in Charlotte, or pass through there to where they need to go. A message may go from Fort Myers (let's say that's procurement) to Burlington (let's say that's the on site team) – but it passes through Charlotte. Each of the nodes is another aspect of the project – whether people, tasks, items, money, reports, or anything else. The central hub of the project, that plans the route map and operates it, is the project manager. Sure, there are other minor hubs (Philadelphia, Washington) – these are team leaders. Even they, ultimately, end up in Charlotte. Charlotte is Project Central. That's the project manager.


Hear the first single "Who's Been Having You Over"
Libertines and Babyshambles frontman Pete Doherty has announced plans for a new album with a new band. Under the moniker Peter Doherty & the Puta Madres, the group will release their self-titled album on April 26 via Strap Originals/Cargo Records.
The album was recorded over four days at a family home overlooking a fishing village in Normandy. It was produced by Jai Stanley and engineered by Dan Cox.
The first single arrives with "Who's Been Having You Over." Stream it below.
The tracklisting opens with "All at Sea," likely a re-recording of a Libertines track from the 2005 What Became of the Likely Lads EP. Other songs include "Narcissistic Teen Makes First XI," "Someone Else to Be" and "A Fool There Was."
The album is available in a variety of formats and bundles, including a deluxe vinyl version that includes a new documentary. See all available pre-order options here.
The new Peter Doherty & The Puta Madres album - out 26th April!
The Libertines last released an album in 2015 with Anthems for Doomed Youth. Doherty also released a solo album titled Hamburg Demonstrations in 2016.
The Libertines "Heart of the Matter"


text:
800PBX Inc. Launches the Best-Rate Toll Free Number PBX Service
800PBX Inc., a leading provider of small business PBX systems, has announced the launch of its first commercial offering of Toll Free PBX Services starting today.
800 PBX Brings Salesforce.com CRM...
Fremont, CA (PRWEB) September 12, 2006
800PBX Inc. a leading provider of small business PBX systems has announced the launch of its first commercial offering of Toll Free PBX Services starting today.
"We are offering hosted PBX services at 3.5c/minute rate with no double billing. This is a direct result of many negotiations with different carriers! Reliable, good service and low price strategy will help us attain the global leader position in the coming two years!" quotes Manohar Chapalamadugu, CEO of the company. "The panel has been worked out for many years and we believe to have gotten it right making many features possible straight away from the panel" add Raj Kumar, Vice President, Product Development of the company.
The advanced services including Automated Attendant, Business and After Business Hours Greetings, Dial by Name, Auto Call, Call Hunt, Follow Me, Fax In/Out, Custom Music on Hold, Operator Extension, Voice Mail and Call Screening features are provided in the first release. Panel is also provided free of cost to customers to reflect their call preferences real time. The company plans to integrate the audio conferencing services by the end of this quarter.
800PBX's proprietary WebDial feature allows the customers to immediately place a call and get connected to the company's main system at one click from the website. WebDial feature is provided free of cost.
Priced generously at $9.99/Month for entry level businesses to $99.99/Month for Enterprises, 800PBX Inc. will be the best cost effective reliable service available presently.
Free 15 day trial with 75 minutes is available with any plan. For a limited time, 800PBX is offering free Vanity Toll Free and 800 numbers. A voice artist is also available free of cost for recording the company greetings.
800PBX Inc. is a leading provider of Small Business PBX and Audio Conferencing services.
For more details, visit http://www.800pbx.com
press @ 800pbx.com
1-888-800-0 PBX (1-877-800-0739)
800PBX Inc.


Pre-pasted, removable wallpaper created from sections of Ashley Lasseigne's artwork and illustrations.
• PVC-free paper, durable, wipeable, eco-friendly and class A fire rated.
Read more about the artist, Ashley Lasseigne, here.


text:
Celebrate and Restore
in collaboration with East Bay Municipal Utility District
Skyline Gardens Alliance
© 2018 by Skyline Gardens Project. Proudly created with Wix.com
skylinegardens@ebcnps.org
Dear Skyliners,
Last week, we were out in good weather weeding and planting. On Sunday, we had another good group and we weeded and planted along First Outcrop Ridge (just beyond the thinned Eucalyptus grove). Welcome to Steve.
So far, we've planted nearly 200 new native plant starts. In terms of our schedule this year for planting and weeding, we're right on target, as we dodge around these welcome rains.
We'll be out regular days this week: Wednesday at 2 and Sunday at 9:30. The weather looks dicey for Sunday, so we may switch to Saturday afternoon instead. Please let me know if you can make it, and if Saturday afternoon would work as a back-up plan.
Last Sunday, we encountered epic fog; thick valley fog was blowing across the ridge from the Diablo Valley. Here's a shot as we climbed up through the Eucalyptus grove:
We felt like pilgrims, climbing the heights in a mystical land. (thanks to Max for this photo)
In our weeding now, we're coming across a number of annual seedlings, so here's several pictures to help us learn them. All three of these guys have fuzzy leaves, so at first glance, they might be easy to confuse.
Here's Popcorn Flower (Plagiobothrys nothofulvus):
Popcorn flower forms rosettes of pointed leaves. With the right light, the hairs at the edge of the leaves are rust colored. In flower, these look like tiny, white Forget-me-nots. They grow on rocky ridges, and are profuse at Diablo Bend, near the new bench.
Next we have Fiddlenecks (Amsinkia menziesii):
These also grow as rosettes with fuzzy leaves, but the leaves are long and rounded at the tips. Fiddlenecks are plants of meadows and hillsides. They are orange when then bloom, and the flowering stalk is curved at the tip, like the end of a fiddle. Incidentally, both Fiddlnecks and Popcorn flower are in the same Borage plant family, so the similarity is based on a close relationship.
Here's our third seedling, one of the Tarweeds. This is Slender Tarweed (Madia gracillis):
This one also has fuzzy leaves, too, but the leaves grow in a cross-pattern (at right angles), so the shape is very distinctive. When in flower, these bear small, yellow daisies on slender stalks a foot or two high. The foliage has a lovely, lemon-scented musk. (Thanks to Josh for all three seeding photos.)
Besides seedlings, we also found the first stirrings of the buds of our big perennials. Here's the buds of Grey Mule's Ears (Wyethia helenoides), just beginning to wake up:
If all goes well, these will be in glorious full bloom by April 1. Each bud contains all the leaves and flowers that will emerge on that stalk in the coming months. (Thanks to Max for this shot.)
Just like fruit trees, who form their new leaves and flowers in "last year's" buds in the summer, the big perennials do the same. As weather warms in the spring, the buds unfold and we see what they were doing last summer, when the leaves looked dead and brown. If you were able to look inside these buds, you would see the future.
Of course, these are wild plants in wild lands, so anything can happen. Last year as the first shoots on this very plant were rising up, we came out one morning to find the tips all browsed off. It could have been a deer, or a rabbit, but no matter who did it, "Bye-bye flowers," but "Hello tasty bite."
As to this year, we shall see.
Happy Trails,
Back to 2018 Skyline Report