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The Gene Editing Institute is the only research institute of its kind in the nation based within a community health care system. Led by gene editing pioneer, Eric Kmiec, Ph.D., and based in ChristianaCare's Helen F. Graham Cancer Center & Research Institute, the Gene Editing Institute is a worldwide leader in gene editing biomedical research in cancer and other inherited diseases, and the only one working in the same space with oncologists, genetic counselors and patients, bringing translational research from basic science to patient treatment to an entirely new level.
Research Scientist II - Gene Delivery will participate in all aspects of preclinical development work in new, targeted delivery system utilizing CRISPR gene therapy to treat NSCLC. The successful candidate will work on the design and evaluation of gene and cell delivery systems including adeno-associated virus and lipid nano particles in experimental lung cancer models. The Research Scientist is also involved in the development of proof-of-concept and preclinical studies for other solid cancers. The ideal candidate will have advanced expertise in adeno-associated virus biology and at least one area in lipid nano particle delivery, animal models in Non-small cell lung cancers, or CRISPR.
Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related death worldwide and in the United States. There is a high unmet need for new treatment options to treat NSCLC due to the high rate of chemoresistance in NSCLC. Gene Editing Institute at Christiana Care Health System has identified a specific target to reduce the chemoresistance and is currently developing CRISPR gene therapy strategy to treat NSCLC. This platform protocol is also applicable to other cancers. Other responsibilities to include:
Requirements: PhD in cancer biology, molecular biology, cell biology, biochemistry, or a closely related field. Minimum of three to seven years of industry experience. Candidates should also have authored at least two first author publication (either published or in the process of submission) in reputable journals; cancer research experience preferred.


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Bank deposits at the European Central Bank have more than doubled to a five-month high and European institutions are paying more for dollars, signs of concern that turmoil in eurozone bond markets could spread.
The ECB said that banks deposited 104.9 billion euro ($150 billion) overnight on Tuesday, the highest level since early February and up from 49.9 billion euro on Friday. The central bank pays below market rates, implying the jump means that banks are hoarding funds rather than lending to each other.
A number of short-term funding markets were unnerved last week by the US debt ceiling drama but analysts said the jump in deposits pointed to more than that.
"The moves we've seen in the past three days suggest this is nervousness based in the euro markets, not elsewhere," said Simon Smith, chief economist at FXPro.
In a further sign of dislocation in short-term funding markets, the rates European banks must pay for dollars have jumped.
A gauge that measures the spread, or difference, between the price of dollars in three month's time and the rates implied by equivalent US and eurozone interest rates has reached levels last seen in the aftermath of the first Greek bail-out last year, according to analysts at Morgan Stanley .
Banks were this week paying 63 basis points over three-month US interbank market rates, the bank said, up from about 30bp last month - though this is still far below the 200bp-plus peak reached following Lehman Brothers' 2008 collapse.
European banks often issue short-term US commercial paper to get dollar funds but this market has slowed as would-be lenders pulled back amid the negotiations over raising the US debt ceiling. This has forced banks instead to turn to the open currency markets, where the surge in demand has in turn led to a squeeze.
Bankers said it was difficult to gauge the significance of the market moves because of the thin holiday-like trading typical of August.
"The real pressure is going to come in September for many banks. We know they were ahead of the curve in their funding in June, but some could come under stress in the autumn if they're still not getting access," said one senior syndication banker.
Italian bank debt has come under particular pressure as yields on the country's sovereign bonds have reached euro-era highs.
"The wholesale market due to the spreads can't be tapped. On the short (term market) liquidity is there," said Marina Natale, chief financial officer of Unicredit, on Wednesday during a call to discuss the bank's results.
Most Spanish banks and savings banks have been struggling for months to raise money in the wholesale finance markets, and the latest jump in sovereign bond yields has made the prospects for the rest of the year grimmer, according to bankers and analysts in Madrid.
No Spanish private sector lender has sold bonds since a poorly-received €1bn covered bond from Santander, the eurozone's biggest bank by market capitalisation, just over two months ago, according to Analistas Financieros Internacionales, a research group.
Spanish banks have been relying heavily on repurchase transactions through clearing houses such as LCH.Clearnet, where they can raise short-term liquidity by posting government bonds as collateral.

subset: commoncrawl


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The Lacs – Make the Rooster Crow
Labels: Average Joe's Entertainment
Number of Tracks: 1
Total Time: 2:57
Formats: Hot AC, Alternative, Rock, Urban, Dance, College, Hip Hop, Mixshow – Rhythm / Urban, Mixshow – Dance, Rhythmic, Rhythmic Mixshow, Rock – Specialty, Urban – Mainstream, Rhythmic AC, Urban Mix Shows
Available Date & Time: May 21, 2015 03:00 PM ET
Country: USA
Drop It Down Low and make The Rooster Crow!
From the new album, Outlaw In Me, coming May 26.
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Process and Procedures for the Student Uniform Referrals
The School Uniform Referral program has improved the distribution procedures to families in need of this assistance. To better serve our community of schools we have created a referral and distribution procedure that better meets the needs of our families, keeping in mind that our school personnel is better suited to initiate the referral for the families in their schools.
We kindly ask that school personnel keep in mind that our non-profit partner Shoes and Clothes for Kids have a limited number of uniforms and these are available for those families that have demonstrated true need. We will be able to provide for our students in grades Pre-K through 8, two uniforms for the child in need (and a set of regular clothes, also included will be a shoe gift card. Please be reminded that this is a one-time gift per school year. For High School students uniform sizes in stock, we will provide 2 sets of uniforms.
- Referrals are made only by CMSD staff, not parents, guardians or community partners; with the exception of Community Wraparound Site Coordinators.
- School Administrator must assign a staff person to coordinate uniform referrals for their school.
- STOP Before placing referral please check with your school Uniform Referral Coordinator to avoid duplicate referrals. This slows the process and delivery.
- Go to CMSD website (ClevelandMetroSchools.org).
- Click on the Staff tab located at the top portion of the District website.
- Under Quick Links go to Uniform Assistance Referral tab on the right hand side of the page and click.
- Complete form. Please refer to the sizing chart when completing this form.
- Click on "Done" when finished. Reminder: be sure and complete the entire referral form, so that we can assure accurate packing of uniforms.
- Please be aware that uniform supplies are limited by funding constraints and some referrals may or not be able to be delivered and depends on uniform availability of our non-profit partner. If possible, we will try and locate gently used uniforms for the student.
- Once referral has been done by 12 noon on Monday, distribution will be set-up for the following week.
- The office of Attendance will update distribution roster that is viewed by Uniform Referral Coordinator at the school.
- Delivery will be done by Guaranteed Delivery Service who also does our "inter-office mail" delivery unless there is an unusual amount of uniforms requested. If this is the case, we will assign a different form of delivery through our internal delivery system. The signature they collect will suffice as proof-of-delivery and be made available to SC4K, as needed.
- In order to deliver the bag of clothing to the student, the Uniform Referral Coordinator must meet with the student's parent/guardian to discuss the need for school attendance, signing of the Attendance Pledge and filling out of the short survey. Documents will then be emailed or sent through inter-office mail to the Office of Attendance. This documentation is critical for funding purposes of this program.
- The assigned school staff will be responsible for sending the Attendance Pledge and survey back to the Office of Attendance.
If you have further questions please contact:
Shoes and Clothes for Kids: firstname.lastname@example.com
Or Office of Attendance:
Victoria Harris
email@example.com /(216)838-0283Your referral will be processed in the order that it has been received. Remember that quantities of uniforms (including sizes) are limited; therefore your referrals must be based on a critical need and distribution will continue as long as supplies last. Please take a few moments to complete the referral form below in order to expedite uniforms for the student in need of this service. It is important that you refer to the size charts below for optimal fit.Student Uniform Referral Form please click here.


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Why is the Child Security of Your Doors and Windows Important?
We come across hundreds of tragic 'kid falls out of window' stories, all of them results of a moment's inattention.
We also come across 'hero passersby catch kid falling out of window' stories, yet these are rare.
We could easily prevent these stories with window security solutions.
TGP Safety Systems for your Children and Home
TGP Window Safety Systems are simple and effective child safety and accident prevention solutions designed to set your mind at ease. TGP Safety Locks keeps the kids inside and the burglars out.
TGP window safety solutions are ideal for any floor of any type of building. Our products help create a safe, accident-free environment and help keep intruders out.


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SachajuanThickening Shampoo
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What Else You Need to Know: This shampoo offers weightless moisture and no residue to support the hair fiber for volume and stability. Its formulation is ideal for fine, limp hair.
-Massage a small amount into wet hair.
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-Heat and UV Protectant: Protects from sun, heat, and external elements.
Aqua, Sodium Laureth Sulfate, Glycereth-2 Cocoate, Cocamidopropyl Betaine, Glycerin, Peg-4 Rapeseedamide, Perfume, Sodium Chloride, Sodium Benzoate, Hydroxypropyl Guar, Hydroxypropyltrimonium Chloride, Citric Acid, Trisodium Sulfosuccinate, VP/DMAPA Acrylates Copolymer, Limonene, Linalool, Benzyl Salicylate, Hexyl Cinnamal, Alpha-isomethyl Ionone, Butlyphenyl Methylpropionbal, Hydrolyzed Rhododphycea Extract, Benzalkonium Chloride, Phenoxyethanol.
Founded in Stockholm, Sweden by close friends Sacha Mitic and Juan Rosenlind, SACHAJUAN Professional Haircare Products has been taking the world by storm since 1997. Esteemed international hairdressers, Sacha and Juan are true craftsmen with a love for their art <i>and</i> their clients—plus an exquisite styling portfolio filled with high-fashion work. With products based largely on the principles specific and unique to their incredibly nourishing Ocean Silk technology, SACHAJUAN is designed to create healthy, strong hair that's radiant and lustrous. Their simple and straightforward formulas blend serious technological innovation with luxury for results that are immediate, unparalleled, and unbelievably beautiful.
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This study aimed at elucidating the physiological basis of bacterial antibiotic tolerance. By use of a combined phenotypic and gene knockout approach, exogenous nutrient composition was identified as a crucial environmental factor which could mediate progressive development of tolerance with markedly varied drug specificity and sustainability. Deprivation of amino acids was a prerequisite for tolerance formation, conferring condition-specific phenotypes against inhibitors of cell wall synthesis and DNA replication (ampicillin and ofloxacin, respectively), according to the relative abundances of ammonium salts, phosphate, and nucleobases. Upon further depletion of glucose, this variable phase consistently evolved into a sustainable mode, along with enhanced capacity to withstand the effect of the protein synthesis inhibitor gentamicin. Nevertheless, all phenotypes produced during spontaneous nutrient depletion lacked the sustainable, multidrug-tolerant features exhibited by the stationary-phase population and were attributed to complex interaction between starvation-mediated metabolic and stress protection responses on the basis of the following reasons: (i) the nutrition-dependent tolerance characteristics observed suggested that adaptive biosynthetic mechanisms could suppress but not fully avert tolerance under transient starvation conditions; (ii) formation of specific phenotypes could be inhibited by suppressing protein synthesis prior to nutrient depletion; (iii) bacteriostatic drugs produced only weak tolerance in the absence of starvation signals; and (iv) the attenuation of the stringent and SOS responses, as well as the functionality of other putative tolerance determinants, including rpoS, hipA, glpD, and phoU, could alter the induction requirement and drug specificity of the resultant phenotypes. These data reveal the common physiological grounds characteristic of starvation responses and the onset of antibiotic tolerance in bacteria.
Bacteria constantly harbor a subpopulation refractory to killing by bactericidal drugs (2, 3, 16). The size of this tolerant subpopulation increases in adverse growth conditions such as depletion of nutrients (6, 11, 32). The physiological basis of this phenomenon, termed antibiotic tolerance, remains poorly understood. Using a systematic approach to screen knockout mutants of Escherichia coli, Hansen et al. recently identified a number of genetic loci involved in the upstream control of a vast tolerance response network (8), suggesting that antibiotic tolerance was not simply due to passive shutdown of drug target activities as a result of retarded bacterial growth in an unfavorable environment. In Hansen's study, most of the genes for which deletion had a detectable effect on antibiotic tolerance were global regulators of nutrient metabolism, stress sensing, and protection as well as essential cellular processes such as transcription. We envision, however, that genetic determinants responsible for mediating response to specific stress may not be readily identified by deletion studies if the test population is subjected to exposure to multiple induction factors, since bacteria may sense alternative stress signals and develop tolerance via redundant pathways. Hence, any attempt to screen for tolerance mutants under uncontrolled conditions, such as those involving stationary-phase culture in which a number of growth parameters become unfavorable, may allow identification of only the rather upstream regulators that control a number of independent tolerance mechanisms or some common downstream protection mechanisms activated by different induction pathways. Indeed, this idea is consistent with the observation that only a small number of highly redundant and upstream regulatory genes were identified through screening of stationary-phase cultures of gene knockout and other mutant libraries (8, 17).
To overcome the limitation of conventional tolerance assays using a stationary-phase population and test our concept on the intermingling network of stress sensing and protection, we strived to dissect and depict the relative roles of the major branches of such network in tolerance development through analysis of the differential effects of progressive nutrient exhaustion, considering that starvation represented a major stress that bacteria encountered during stationary phase. In this work, a tolerance induction assay was designed in such a way that the test conditions were tightly controlled to evaluate the induction effect of a panel of nutrient compositions. Through a series of conditioned experiments, we were able to identify the key nutritional factors and determine their relative roles in tolerance induction and demonstrated that starvation of each nutrient category rapidly elicited matching metabolic and physiological changes which might in turn be reflected by phenotypic tolerance to antibiotics.
On the basis of the bacterial tolerance induction criteria gathered in the phenotypic experiments, we further attempted to perform a preliminary functional mapping of several major bacterial stress defense mechanisms in mediating formation of the nutrient-sensitive antibiotic tolerance phenotypes. These include the stringent (19, 26, 27), SOS (10, 21), and RpoS (13) mediated responses, which may be elicited when bacteria encounter nutrient starvation, physiological stress, or cellular damages as well as several other key genetic loci, such as hipA (12, 22), phoU (17), and glpD (30), which have been implicated in the formation of bacterial persister subpopulations that exhibit markedly reduced death rates during antimicrobial treatment. Currently, the relative effects of the depletion of different nutrient classes on the induction of these stress responses remain unclear. For example, it has previously been shown that starvation of glucose, amino acids, and ammonium salts could each result in differential expression or stability of the RpoS factor (9, 25), which is known to regulate the expression of a range of protection mechanisms, such as thermotolerance (18) and oxidative stress defenses (20); however, it is not clear whether different types of nutritional stress can activate such RpoS-controlled mechanisms, as well as the stringent and SOS responses, on an equal basis. Apart from a lack of information on the induction criteria of different tolerance responses, the pattern of interaction between these systems remains poorly defined. For example, the stringent response is known to enhance the activity of the RpoS sigma factor (9, 19), yet information regarding whether stringent response can also elicit the SOS response is not available. On the other hand, a number of recent studies suggested that bactericidal antibiotics could trigger the production of deleterious hydroxyl free radicals by destabilizing membrane integrity and eliciting the onset of Fenton reactions (14, 15). In view of the wide range of stress responses which can regulate protein translation efficiency as well as production of antioxidant defense actions, we hypothesize that formation of antibiotic tolerance during nutrient depletion is at least partly due to the fact that reactive oxygen species produced during the process of drug-induced physiological changes are neutralized by the protection effects of stress responses elicited under nutrient starvation. Upon testing specific deletion mutants against a range of conditions which were known to produce antibiotic tolerance, we were able to assess the relative role of each stress response component and the redundant nature of the key systems involved in tolerance formation. Our data showed that deletion of a specific gene often resulted in condition-dependent variations in tolerance phenotypes. Such data confirm the presence of a complex, redundant, and hitherto undefined nutrient-sensing and tolerance response network which can regulate bacterial physiological changes and the concomitant formation of drug-specific tolerance phenotypes in accordance with the nutritional status in the environment.
Bacterial strains and culture media.The Escherichia coli K-12 strains and derivatives thereof listed in Table 1 were used in all experiments. These strains included a ΔrelA ΔspoT double knockout mutant and the wild-type strain from which this mutant was constructed (30) as well as deletion mutants of specific genes derived from the Keio collection of E. coli knockout strains (1). All cultures utilized rich defined medium (RDM) (Teknova, Holister, CA) unless stated otherwise. This medium, developed by Neidhardt et al. (23) and later modified by F. R. Blatter and coworkers (http://www.genome.wisc.edu/ ), comprised six major components: (i) MOPS (morpholinepropanesulfonic acid) base, (ii) glucose, (iii) ammonium salts, (iv) inorganic phosphate, (v) nucleobases, and (vi) an amino acid mixture with trace vitamins. The composition of this growth medium could be precisely manipulated according to the manufacturer's instructions (http://www.genome.wisc.edu/resources/protocols/ezmedium.htm ). On the basis of the RDM recipe, a panel of defined media consisting of different compositions as denoted in the present study was constructed for the tolerance induction assays. Standard LB agar (Difco, Leeuwarden, The Netherlands) was used for assessing the proportion of the bacterial population that survived in the assay.
Drugs and chemicals.All antibiotics were purchased from Sigma (St. Louis, MO). Sodium azide and potassium cyanide were obtained from Merck and BDH Chemicals, respectively.
Growth and assay media.Fresh bacterial colonies were inoculated into RDM and grown overnight at 37°C under constant agitation (200 rpm). The overnight culture was diluted 10,000-fold in RDM and cultivated for 4 h until the exponential growth phase was reached (∼107 CFU/ml). Aliquots of this exponential-phase culture were washed and resuspended in the assay medium, followed by the tolerance induction assays.
(i) For analysis of the effect of different nutrient compositions on induction and maintenance of tolerance to specific bactericidal antibiotics, various recipes, ranging from complete RDM to one which was deprived of all test nutrients (MOPS), were prepared (Fig. 1). Specific growth rates (μ) prior to antibiotic treatments were determined by measuring the changes in cell density in the untreated controls, using the formula μ = [(log10N3 − log10N0) 2.303]/(t3 − t0), where N0 and N3 represent the initial and final cell densities, respectively, within the first 3 h of incubation upon the switch to the test medium (t3 − t0).
(ii) For comparative assessment of the tolerance induction effect of bacteriostatic conditions in a nutrient-rich environment, complete RDM was supplemented with individual bacteriostatic agents (tetracycline [2 μg/ml], rifampin [16 μg/ml], 5 mM sodium azide, and 5 mM potassium cyanide) and used to resuspend the test population prior to the assays.
(iii) To determine whether the multidrug tolerance phenotypes observable in stationary-phase bacterial culture were attributable to diminished nutrient availability, filter-sterilized cell-free supernatant from RDM-grown overnight cultures was prepared and tested for its tolerance induction effect in tolerance induction assays.
(iv) For genetic mapping of tolerance induction pathways, a panel of selected deletion mutants was tested against specific recipes of nutrient compositions to determine whether deletion of specific genes would affect the ability by which bacteria produced specific tolerance phenotypes under conditions which were known to induce tolerance formation. The gene knockout mutants and the corresponding tolerance induction conditions tested are shown in Fig. 5 and 6. Potential development of suppressor mutants from the guanosine 3′,5′-bispyrophosphate (ppGpp)-null mutant was assessed by plating out all test populations of this strain on M9-salt agar (29) supplemented with 0.4% (wt/vol) glucose.
Tolerance induction assay.Bacterial populations were preincubated with the test medium at 37°C for 2 h and subjected to antibiotic challenge using ampicillin (100 μg/ml), ofloxacin (0.75 μg/ml), and gentamicin (6.25 μg/ml) at a working concentration of 25 times the respective MIC of the test strain (BW25113). Cells were incubated for 48 h at 37°C under constant shaking (200 rpm). Standard serial dilution and plating on LB agar was performed at 0, 3, and 48 h. The plates were incubated at 37°C, and colony counts were recorded for two consecutive days to account for possible discrepancies due to residual drug effects. Changes in the sizes of surviving cell populations over time were recorded and compared to the level observed prior to antibiotic challenge for assessment of short-term (3-h) and long-term (48-h) drug tolerance. At least three independent experiments were performed in each assay to assess the reproducibility of the induction effect. Surviving cells were routinely tested for antibiotic susceptibility by using the agar dilution method to determine the MICs of isolates randomly selected from the tolerant population.
Tolerance inhibition assay.To test the effect of transcription and protein synthesis inhibitors on starvation-induced tolerance development, tetracycline (4 μg/ml) was added to an RDM-grown log-phase culture and incubated for 2 h at 37°C, and the treated population was subjected to centrifugation, washed once, resuspended in MOPS base which also contained tetracycline at 4 μg/ml, and incubated at 37°C for 2 h. The cell suspension was then subjected to antibiotic challenge as described for the tolerance induction assay.
This study involved the development of a phenotypic assay to probe the induction criteria and physiological mechanisms governing bacterial antibiotic tolerance formation. In the assay, a panel of specially formulated media with known nutrient compositions was used to analyze the effects of limitation of different combinations of five key nutrients (amino acids, glucose, ammonium salts, phosphate, and nucleobases) in eliciting tolerance development in an actively growing bacterial population which was fully sensitive to the bactericidal effects of the three test drugs (ampicillin, ofloxacin, and gentamicin) prior to the assay. Figure 1 shows the relative survival rates of the test population recorded under each assay condition before and after drug treatment for 3 and 48 h and thereby the relative capacity of a bacterial population to develop and maintain tolerance to a specific drug under each assay condition. The ability of the test population to maintain viability or propagate in the assay medium prior to or without drug treatment, illustrated as specific growth rate and alteration in population size before and after treatment, respectively, was also shown to depict the relationship between the growth capacity and the corresponding drug susceptibility profiles of the test organisms. In all assays, the bacterial population size was standardized to approximately 107 cells per ml prior to drug treatment.
Four major classes of drug tolerance responses.The diverse spectrum of nutrient compositions tested was found to produce complex and condition-dependent antibiotic susceptibility phenotypes which could be divided into four major categories according to their drug specificity and sustainability (Fig. 1). First, a total of five conditions, including the full RDM recipe, failed to support survival against the bactericidal effects of the three test drugs (Fig. 1, category A). These conditions mainly involved depletion of a single nutrient except amino acids but also included one in which three nutrient classes (glucose, ammonium salts, and nucleobases) were simultaneously depleted from the growth medium. Second, two conditions could specifically produce transient tolerance to ofloxacin but not to the other two drugs, and in both cases, phosphate was depleted from the medium (Fig. 1, category B). Third, a total of 11 recipes produced mixed tolerance responses to ampicillin and ofloxacin (Fig. 1, category C). The recipes that produced category C responses ranged from a lack of amino acids alone (Fig. 1, C1iii) to simultaneous depletion of up to four nutrients (Fig. 1, C3). Apart from specificity and sustainability, the strength of drug tolerance, recorded as the size of the tolerant population produced and maintained during the assay, also differed extensively between different nutrient compositions. Finally, eight conditions, all involving media which lacked glucose and amino acids, were found to elicit tolerance to all the three test drugs. The phenotypes that belonged to this category typically included sustainable tolerance to both ampicillin and ofloxacin and transient tolerance to gentamicin (Fig. 1, category D).
Inductive and regulatory effects of amino acids and nucleobases.Detailed analysis of the relationship between nutrient compositions and the corresponding drug susceptibility phenotypes revealed several important tolerance induction characteristics which could be made apparent only by comparing the induction effects of differential nutrient compositions. First, we noticed that depletion of amino acids from the growth medium was a prerequisite for the onset of the vast majority of tolerance phenotypes, with the exception of several phosphate-limiting recipes which could induce short-term tolerance to ofloxacin or to ofloxacin and ampicillin even in the presence of amino acids (Fig. 1, Bi and -ii and C1ii, -vi, and -vii). Hence, depletion of amino acids produced a much more diverse and far-reaching effect than the lack of any other compounds in eliciting antibiotic tolerance, although the sustainability and drug specificity of the phenotypes produced were highly dependent on the nature of the nutrients which were concomitantly depleted.
The second important feature observable during nutrient depletion-mediated tolerance induction is that several induction processes appeared to be highly sensitive to the relative abundances of nucleobases. In the case of ampicillin but not ofloxacin, simultaneous depletion of amino acids and nucleobases consistently produced a much weaker induction effect than a medium which was deprived of amino acids only (Fig. 1, compare C1i and C1iii). Although this finding appeared to indicate that the presence of nucleobases could enhance the development of ampicillin tolerance mediated by depletion of amino acids, a contradictory role for these molecules in tolerance regulation was observable in assays with other nutrient compositions. Notably, a prolonged ampicillin tolerance phenotype conferred by a lack of amino acids and ammonium salts could not be supported if nucleobases were present (Fig. 1, compare C1iv to C2ii), suggesting a suppressive effect of nucleobases on specific tolerance induction pathways. In contrast, prolonged ampicillin tolerance triggered by glucose limitation was not affected, as long as amino acids were absent (Fig. 1, Di and -iv). Apart from ampicillin tolerance, the specific inhibitory effects of nucleobases were also observable in ofloxacin tolerance induction; in this case, simultaneous depletion of amino acids and phosphate produced sustainable ofloxacin tolerance only in the absence of nucleobases (Fig. 1, compare C1v to C4i).
Induction of prolonged tolerance by depletion of multiple nutrients.The third important feature of nutrient-sensitive tolerance regulation is that bacteria were able to exhibit prolonged tolerance to antibiotics under specific conditions in which key nutrients were further removed from an amino acid limitation background. This finding was mainly based on our observation that highly sustainable tolerance of ampicillin and/or ofloxacin could be produced only by simultaneous depletion of amino acids and glucose (Fig. 1, category D), or amino acids and ammonium salts, plus either phosphate or nucleobases (Fig. 1, C2, -3, and -4). Absence of amino acids and glucose was also a prerequisite for induction of short-term tolerance to gentamicin, a phenotype which consistently emerged along with sustainable tolerance to ampicillin and ofloxacin (Fig. 1, category D).
Our data indicated that prolonged antibiotic tolerance was mediated by complex adaptive and stress responses inducible when specific nutrients other than amino acids became limiting. Such responses, apart from being subjected to negative regulation by amino acids, appeared to produce induction effects synergistic with those of amino acid depletion. First, we noted that simultaneous limitation of glucose, ammonium salts, and phosphate did not support even short-term ampicillin tolerance when amino acids were present (Fig. 1, Bii and C1ii), yet the presence of these three nutrients could reverse most of the MOPS-induced phenotypes even in the absence of amino acids (Fig. 1, compare C1i with category D conditions). We therefore hypothesized that a lack of one or more of these compounds could induce a tolerance response which was negatively regulated by amino acids and distinguishable from the one triggered by amino acid depletion alone, which was nucleobase dependent (Fig. 1, compare C1i to C1iii). We noticed that, although each of these components could not fully reverse MOPS-induced ampicillin tolerance (Fig. 1, C3i and Dvi and -vii), further depletion of ammonium salts or glucose (Fig. 1, C2ii and Div), but not phosphate (Fig. 1, C4i), from a medium background which failed to induce the putative amino acid depletion-mediated ampicillin tolerance response (lack of amino acids and nucleobases simultaneously) (Fig. 1, C1i) could induce sustainable ampicillin tolerance. Corroborating the idea that prolonged tolerance was regulated by derepressible mechanisms was the observation that ofloxacin tolerance was inducible by two independent pathways. First, a lack of either amino acids or phosphate induced primary tolerance to ofloxacin after 3 h of treatment (Fig. 1, Bi and C1iii), the phosphate depletion-mediated response being insensitive to suppression by amino acids. Second, a stronger and more persistent phenotype resulted from a combined depletion of amino acids, phosphate and nucleobases, or amino acids and carbon sources (Fig. 1, C3i, C4i, and all category D conditions). It should also be noted that, unlike amino acids and nucleobases, glucose, phosphate, and ammonium could not individually confer negative regulation on antibiotic tolerance formation. In this study, we confirmed that long-term tolerance to ampicillin and ofloxacin was not due to selection and proliferation of resistant mutants, since all tolerant populations were found to contain isolates which remained susceptible to the test drugs in MIC determination.
Phenotypic difference between starvation- and stationary-phase-induced tolerances.To assess the relative contribution of nutrient deprivation in producing multidrug tolerance in stationary-phase populations, we performed a spent-medium assay to determine if the cell-free supernatant obtained from stationary-phase culture, which had been grown overnight in RDM, could induce formation of a phenotype which matched those produced by the tested recipes. Our results showed that this spent medium supported tolerance development in a manner similar to that of MOPS base (Fig. 2). We showed that this induction effect was due to nutrient limitation but not accumulation of toxic metabolites produced during prolonged culture, as it could be abolished by replenishment of nutrients to the spent medium (results not shown). These findings confirmed that nutrients in an overnight culture have been depleted to a level that would support development of tolerance phenotypes comparable but not entirely similar to those observed in stationary-phase populations, which exhibited prolonged gentamicin tolerance.
Growth rate and antibiotic tolerance.In addition to the relationship between nutrition recipes and tolerance formation, we further analyzed the correlation between growth rate and phenotypic tolerance under the test conditions. We found that a high specific growth rate (an average of 1 h−1 or above between 0 and 3 h) (Fig. 1, category A) was not compatible with tolerance development, whereas conditions that suppressed bacterial growth (with specific growth rates of 0 h−1 or below) generally conferred sustainable tolerance to both ampicillin and ofloxacin as well as short-term tolerance to gentamicin (Fig. 1, Di to -iv and -viii). However, the relationship between growth rate and drug susceptibility became much less apparent under conditions in which bacteria grew at a relatively slow pace, when induction of antibiotic tolerance in a growth rate-independent manner was often observed. For example, a medium which was depleted of amino acids and ammonium salts induced significant short-term tolerance to ampicillin (Fig. 1, C1iv), yet another medium, which lacked glucose, ammonium salts, and nucleobases (Fig. 1, Av), did not confer detectable ampicillin tolerance, despite the fact that the test populations exhibited similar growth rates in these media (exhibiting specific growth rates of 0.69 h−1 and 0.62 h−1, respectively). Likewise, a number of other conditions that supported identical growth rates (ranging from 0.32 to 0.36 h−1) were found to induce a spectrum of tolerance phenotypes with significant variation in drug specificity and degree of sustainability (Fig. 1, C1v, C4i, and Dv). In addition, we do not observe significant correlation between the capacity to exhibit prolonged growth or regrowth in the test medium (exhibited by increase in population size between 3 and 48 h) (Fig. 1), which is a hallmark of adaptive biosynthesis, and development of sustainable tolerance.
Tolerance induction potential of bacteriostatic agents.To determine whether growth inhibition alone could contribute to tolerance formation, we examined the effects of bacteriostatic agents in eliciting tolerance in an actively growing population. Potassium cyanide and sodium azide, both inhibitors of aerobic respiration, were the only agents which continued to exert bacteriostatic effects throughout 48 h of treatment (Fig. 3). In general, the tolerance profiles elicited by bacteriostatic agents were highly dissimilar to those conferred by MOPS or limitation of major nutrients, which consistently supported prolonged tolerance to ampicillin and ofloxacin and short-term tolerance to gentamicin (Fig. 1, category D). Potassium cyanide was the only agent which produced prolonged tolerance to both ampicillin and ofloxacin and elicited short-term gentamicin tolerance in a situation where starvation signals and detectable bacterial growth were both absent (Fig. 3), although the tolerant population was smaller than that observed under nutrient depletion conditions. Interestingly, the bacteriostatic effects triggered by sodium azide were not associated with any detectable tolerance phenotypes (Fig. 3).
Effect of translation inhibitor on MOPS-induced tolerance.We further tested the possibility that active cellular mechanisms might be involved in tolerance formation during nutrient starvation by assessing the effects of tetracycline, a protein synthesis inhibitor, on the potential of MOPS base to induce and maintain tolerance upon corruption of the bacterial protein translation machinery. To avoid potential development of tolerance to tetracycline itself, which would otherwise prevent us from accurately assessing its effects on tolerance to other drugs, we added tetracycline to RDM-grown log-phase culture and continued incubation at 37°C for 2 h prior to switching the culture medium to a MOPS base which also contained the agent, followed by treatment with the three test drugs and assessment of population survival. As shown in Fig. 4, the presence of tetracycline was found to confer a significant but incomplete inhibitory effect on MOPS-induced long-term tolerance to ampicillin as well as short-term tolerance to gentamicin. However, formation of ofloxacin tolerance was not affected by prior inhibition of protein synthesis. It should be noted that this inhibitory effect was not observed when tetracycline was added only after the switch to MOPS instead of 2 h prior to the switch (results not shown), indicating that bacteria might also develop tolerance to tetracycline during starvation. In view of this finding, we speculated that the inhibitory effect of this protein synthesis inhibitor might have been suppressed when the cells became tolerant; hence, the detrimental effect of corrupting protein synthesis on MOPS-induced tolerance development might have been underestimated.
Identification of tolerance response determinants.In a preliminary attempt to identify the genetic determinants responsible for induction of the specific tolerance phenotypes that we observed in the tolerance induction assays, we selected a panel of putative tolerance genes which had previously been implicated in tolerance formation and tested whether deletion of these genetic components would undermine the ability of bacteria to develop antibiotic tolerance under conditions known to induce formation of differential phenotypes. In particular, we included a ΔrelA ΔspoT double knockout mutant, which was defective in producing the stringent response, to determine if the tolerance induction potential of amino acid depletion and the regulatory effects of nucleobases on tolerance formation were mediated through this major starvation response pathway. As shown in Fig. 5 and 6, all the six gene knockout mutants tested displayed tolerance phenotypes which differed from each other as well as from that of the wild-type strain in terms of induction criteria, drug specificity, and the size of the resultant tolerant population that emerged under specific test conditions. In particular, the ΔrecA mutant failed to develop ofloxacin tolerance under all conditions, including stationary phase, yet production of ampicillin and gentamicin tolerance was generally not affected, except under specific conditions, such as the use of overnight culture or a medium in which amino acids alone were depleted, where development of sustainable gentamicin tolerance and transient ampicillin tolerance, respectively, was affected (Fig. 6).
The ΔrelA ΔspoT double knockout mutant, which was defective in producing the stringent response, exhibited pleiotropic effects on tolerance formation. First, this mutant displayed a reduced capacity to produce sustainable ofloxacin tolerance under two conditions (involving MOPS and RDM minus amino acids and nucleobases) which were known to induce prolonged tolerance to this drug in its wild-type counterpart (CF1943) (Fig. 5). Second, the phenomenon of nucleobase-dependent induction of ampicillin tolerance was not observable in this mutant background, as the proportions of the test population that became tolerant to ampicillin were similar regardless of availability of nucleobases in the growth medium. Third, the level of prolonged ampicillin tolerance was reduced in a medium in which both amino acids and glucose were simultaneously depleted. However, such effect was not observable in MOPS base. It should also be noted that, in the absence of antibiotics, the survival rate of this mutant was significantly lower than that of the wild-type strains when nucleobases were supplemented in an amino acid-depleted medium (Fig. 5). However, this mutant exhibited prominent growth when nucleobases were absent in the amino acid depletion medium. On the other hand, the ΔrelA ΔspoT mutant was not capable of surviving for 48 h in a medium depleted of amino acids and ammonium salts; hence, we were not able to examine if the suppressive effect of nucleobases on the ampicillin tolerance phenotypes inducible by a lack of amino acids and ammonium salts was observable in the absence of stringent response. Nevertheless, the ΔrelA ΔspoT mutant was capable of expressing short-term ampicillin tolerance in this medium (results not shown). We confirmed that the test populations of this strain did not harbor RNA polymerase suppressor mutants under all treatment conditions, as no viable colonies were recoverable in minimal glucose agar; hence, all phenotypic characteristics of this strain could be attributed to the lack of ppGpp production.
Apart from observations in the ΔrecA and ΔrelA ΔspoT mutants, deletion of the rpoS gene was also found to consistently result in a slight defect in formation of ampicillin tolerance (Fig. 6). However, development of ofloxacin and gentamicin tolerance was not affected in the ΔrpoS mutant. Finally, deletion of several other genes which were suggested to be key regulators of antibiotic tolerance, including phoU, glpD, and hipA, was also found to produce detectable effects on antibiotic tolerance induction in a condition- and drug-specific manner (Fig. 6).
Overview of tolerance regulatory network.This study established a functional link between bacterial starvation responses and development of antibiotic tolerance. Our data suggested the existence of complex cellular mechanisms that regulated bacterial physiology in accordance with the nutritional status in the environment and effectively exerted sensitive control over the strength and specificity of antibiotic tolerance induction. The major regulatory strategies are summarized as follows. First, deprivation of amino acids, alone or combined with limitation of other compounds, can trigger both primary and synergistic tolerance responses, producing an optimal ratio of actively growing and dormant populations which best enhances population survival under the prevailing environmental condition; second, nucleobases confer either positive or negative regulatory effects on the development of the basal phenotypes, depending on the triggering factors, thus fine-tuning the core responses; third, several nutrient classes, including carbon and nitrogen sources, either individually or concomitantly with each other, can selectively suppress tolerance triggered by depletion of other nutrients, thereby imposing sophisticated negative control on tolerance development. Amino acids are nevertheless the key regulator, as their presence inhibits formation of most phenotypes. The differential, interdependent, counterinteractive, and summative features of tolerance induction indicate that bacteria rely heavily on nutrient-sensing pathways to activate matching metabolic adaptation and stress protection mechanisms, which in turn produce differential antibiotic tolerance phenotypes.
Effect of adaptive metabolism.Our phenotypic data reflect to some extent the result of interplay between the multiple physiological effects exerted by changing levels and composition of endogenous intermediate metabolites during the dynamic process of bacterial adaptation to transient starvation of different nutrients. Such interactive effects on growth rate, drug target metabolism, and production of signaling molecules that regulate stress responses render it impossible to determine the exact role of starvation-induced adaptive mechanisms in tolerance formation. Nevertheless, analysis of the differential phenotypes concerning the availability of nucleobases suggests that they may play a role in enhancing the mRNA synthetic ability and thereby the active production of stress protection proteins, which have been compromised as a result of repression and slow induction of purine and pyrimidine nucleotide biosynthesis under various test conditions (24). In fact, the intrinsic ability of the test organisms to derepress biosynthetic pathways that replenish the depleted ingredients often fails to compensate for the induction effects elicited by the lack of specific nutrients, especially under amino acid depletion conditions. It is envisaged that the regulatory effects of adaptive metabolism in tolerance development diminish upon prolonged starvation, giving way to active stress defense which then plays a key role in conferring sustainable tolerance.
Relative role of stringent response in tolerance development.Being one of the key cellular systems that mediate stress protection in bacteria and one that is activated in an amino acid-limiting environment through induced synthesis of the alarmone guanosine 3′,5′-bispyrophosphate (ppGpp) (26), the stringent response may best account for the nutrient-dependent drug susceptibility phenotypes observable in this study. Given its inhibitory effects on the synthesis of macromolecules such as DNA, phospholipids, and cell wall peptidoglycan (19, 31), initiation of the stringent response in the absence of amino acids is therefore highly consistent with the onset of ampicillin and ofloxacin tolerance under such conditions. This idea may further explain the observation that nucleobases were required for elicitation of amino acids starvation-induced ampicillin tolerance (Fig. 1, C1i and -iii). The results of our gene knockout experiments, however, suggest an alternative story, because deletion of the genetic determinants responsible for producing the stringent response had mixed effects on the differential tolerance phenotypes observed in the wild-type strains (Fig. 5). First, ofloxacin tolerance was not sustainable in this mutant, suggesting that induction of the stringent response was nevertheless required for formation of tolerance to fluoroquinolones. The fact that tolerance to ampicillin and gentamicin could still be produced in the absence of stringent response, however, suggested that its effect was drug target specific. Second, the nucleobase dependency in formation of transient ampicillin tolerance when amino acids were depleted from the growth medium was relieved in the ppGpp-null background, suggesting either that stringent response did not play an important role in formation of ampicillin tolerance under amino acid-limiting conditions or that failure to produce stringent response resulted in derepression of an unidentified mechanism which could then mediate formation of primary or transient tolerance to ampicillin in the absence of both amino acids and nucleobases. These results complement those reported by Hansen et al. (8), which showed that deletion of dksA, a determinant of a ppGpp-dependent modulator of RNA polymerase, resulted in a significant defect in tolerance formation.
The ppGpp-null mutant exhibited enhanced growth fitness in an amino acid-limiting background if nucleobases were further depleted (Fig. 5). It has previously been suggested that a lack of stringent response would undermine the ability of a bacterial population to switch to a defense mode at the expense of continued synthesis of macromolecules during nutrient starvation, thereby enabling the ppGpp-null mutant to grow at a normal or even enhanced rate under transient nutrient-limiting conditions (7, 26). Evidence gathered in this study shows that nucleobases have a significant negative impact on bacterial growth fitness under specific nutritional stress; whether such effect is mediated through complex, stringent-response-independent growth capacity/stress response regulatory pathways entails further investigation. Nevertheless, the intriguing findings regarding the multiple effects of nucleobases on growth rate and tolerance development under different genetic and nutritional backgrounds complement our limited knowledge on the role of ppGpp in eliciting antibiotic tolerance in response to nutrient starvation (27, 28).
Molecular basis of prolonged antibiotic tolerance.In this work, we obtained evidence that depletion of nutrients other than amino acids was involved in expression of secondary responses that triggered onset of prolonged tolerance. Importantly, the tolerance induction effects of glucose-limiting conditions appeared to be suppressed if amino acids were available; once this suppression was lifted, the respective stress responses triggered by depletion of these nutrients could be superimposed on each other, producing a synergistic effect and thereby prolonged tolerance. The stringent response was found to play a role in regulating such secondary mechanisms in a drug-specific manner, as only formation of prolonged tolerance to ofloxacin, but not ampicillin, was affected when the ability to launch stringent response was abolished (Fig. 5). The role of stringent response in development of sustainable antibiotic tolerance was also analyzed in the context of the functional relationship of stringent response with the stationary-phase sigma factor RpoS, which is known to be activated during limitation of carbon and nitrogen sources, as well as phosphate, and to play roles in regulating cellular metabolism during nutrient starvation (25) and in upregulating the transcription of genes involved in stress protection (33). First, carbon starvation was shown to enhance ppGpp production (26), which might in turn activate RpoS-dependent responses (19). Second, our data clearly demonstrated that distinct but overlapping physiological changes were elicited when amino acids and glucose were individually or concomitantly depleted, yet deletions of RpoS and the stringent-response determinant exerted differential effects on the development of prolonged tolerance. These findings indicate that each of these two major stress response systems plays a partial and nonidentical role in formation and maintenance of antibiotic tolerance.
Tolerance induction specificity of SOS response and other key determinants.The finding that RecA function was essential for ofloxacin-specific tolerance was consistent with the role of RecA in mediating DNA repair and therefore alleviation of the damaging effects of fluoroquinolones (5) (Fig. 6). Further investigation is required to determine whether RecA-mediated SOS response plays a role in tolerance formation by promoting DNA repair only or activation of additional protection mechanisms. Currently, there is little evidence which illustrates that SOS response can be spontaneously turned on upon nutrient depletion. Our data suggest a need to examine the possible relationship between the stringent and SOS responses in tolerance formation, as both the ΔrelA ΔspoT and the ΔrecA mutants were defective in exhibiting prolonged tolerance to ofloxacin but not ampicillin (Fig. 5 and 6).
Apart from the major stress responses, the finding that deletion of several putative tolerance determinants which have been postulated to execute a diversity of physiological roles (hipA, glpD, and phoU) (Fig. 6) could in each case affect nutrient-sensitive induction of tolerance in a condition and drug specific manner supports our original hypothesis that the true nature of cellular responses that produced the sustainable phenotypes might involve multiple stress-defense mechanisms regulated by some hitherto un-identified and overlapping signaling pathways. The multiplicity of genetic and nutritional factors observable in bacterial antibiotic tolerance induction confirmed that the underlying regulatory processes were nutrient sensitive, redundant, and additive in nature. The phenotypic difference between starvation-induced tolerance and those observable in an overnight-grown population also suggests that nutrient depletion is not the only tolerance induction factor. Whether the genetic determinants which were found to alter the tolerance strength in a stationary-phase population (8) play a common role in both starvation-specific and nutrient-independent tolerance induction remains to be elucidated.
Concluding remarks.Findings in this work support a tentative conclusion that short-term antibiotic tolerance may develop under amino acid-limiting conditions, which cause reduction in growth rate and activation of stress responses such as ppGpp-related inhibition of drug target function (28, 31), rendering bactericidal drugs ineffective. The inverse relationship between reduced drug target activities and tolerance formation was demonstrated by the effects of bacteriostatic agents, which invariably induced transient tolerance (Fig. 3). The findings that mere inhibition of bacterial growth under specific nutrient-limiting conditions or upon treatment with bacteriostatic agents did not necessarily support tolerance formation are consistent with the idea that cellular integrity could not be maintained for a prolonged period when cell wall synthesis or DNA replication/synthesis was inhibited, unless additional protection mechanisms, such as RecA-mediated target site repair, were also activated. On the other hand, the finding that gentamicin-mediated bactericidal action was not tolerated in the long term implied that the ability to produce starvation-induced tolerance might be compromised without the normal protein synthesis machinery. Such finding is also in agreement with the suppressive effects of bacteriostatic protein synthesis inhibitor, and lack of nucleobases or raw materials for mRNA biosynthesis, on tolerance formation, presumably as a result of reduced capability to launch active protection mechanisms such as antioxidant defense (4) under such conditions. Arguably, the combined effect of nutrient starvation and treatment with protein synthesis inhibitor may impose a dilemma in which bacteria have to balance between the benefits of activation of stress protection mechanisms, which inevitably require active protein synthesis, and stringent-response-related shutdown of the protein synthesis apparatus, which may indirectly alleviate the detrimental effects of mistranslated proteins caused by protein synthesis inhibitors (15). However, the fact that stationary-phase populations exhibit prolonged tolerance to gentamicin (Fig. 2) suggests that bacteria may also possess tolerance strategies that either prevent entry of this drug or circumvent the need to synthesize stress protection proteins. In summary, our data have provided an important framework for further characterization of the molecular basis of bacterial antibiotic tolerance and delineation of the complex signal sensing and protection mechanisms involved in these processes. Such information shall facilitate development of more-effective antimicrobial strategies through assessment of the nutritional impact of in vivo microenvironment on tolerance formation, as well as devising antibacterial stress defense agents that suppress tolerance induction in specific pathogens.
We thank Michael Cashel and Hirotada Mori for providing the stringent-response mutants and the Keio Collection strains, respectively.
- Received 28 August 2009.
- Returned for modification 17 November 2009.
- Accepted 4 January 2010.
- Copyright © 2010 American Society for Microbiology


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Your tour guide will meet you at your hotel at 11:00 - 12:00pm. After about an hour's driving, you will get to the Zhujiajiao Water Town, an ancient Water Town with a history of more than 1700 years and the best-preserved among the four ancient towns in Shanghai. Stroll around the unique old bridges, well-structured courtyards and calm rivers to feel the antiquity, leisure and tranquility of this ancient town. Boat riding along the river is available at your own expense.
Later return to Shanghai downtown and pay a visit to the Shanghai Silk Museum. Afterwards, head to the Bund.
The Bund refers to a waterfront area and is known as the "International Architecture Exhibition Group". Due to its unique geographical location and its influence on the economic activities of Shanghai and China over the past century, it has a very rich cultural connotation. As a symbol of Shanghai, it is a must-see for travelers. Here you will board the Huangpu River Night-view Cruise. Cruising on the river in the evening, you can experience its silence and beauty under the moonlight, enjoy the spectacular sight along the river, and appreciate shanghai dramatic and splendid night view. Afterward, the guide will transfer you back and drop you off at the hotel to conclude the trip.


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Wednesday, March 28, 2012
RIP Earl Scruggs : Foggy Mountain Breakdown
3 comments:
Of that is sad. I knew it was going to happen soon.ReplyDelete
I started playing banjo a few years ago and one the first things I learned was the impact Scruggs had on playing styles. "Scruggs rolls" are repeated sequences of picking using three fingers with the right hand (if you're right handed). It's really that element that gives the banjo it's pickity sound.
Foggy Mountain Breakdown had a mini revival in the chae scene from "Bonnie And Clyde." link.
R.I.P. Earl
Another banjo great to keep an out on (not like a death watch) is Pete Seeger. He's really getting up there in years. Bela Fleck is a youngster by comparison.ReplyDelete
I consider the banjo the quintessential American stringed instrument. Though its origins are African, it was perfected here.
The mere sound of the banjo drives the right people crazy too. I once read that David Crosby forbid the instrument from anything he ever recorded.
I hadn't heard. Sad news indeed. Monster talent in that video.ReplyDelete


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text: News Life Bacteria's amyloids display surprising structure Protein clumps in S. aureus differ from those in Alzheimer's, Parkinson's PROTEIN POWER Proteins from Staphylococcus aureus bacteria make amyloid-like clumps (the streamerlike structures in this illustration) that are toxic to human cells. Dima Abelski/Dimedia Share this:FacebookTwitterPocketRedditPrintEmail By Laurel Hamers February 23, 2017 at 3:00 pm This article is only available to Science News subscribers. Subscribers, enter your e-mail address to access our archives. Client key* E-mail* Log in Not a subscriber? Science News is a nonprofit. Support us by subscribing now. Subscribe


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Many seek the face of a ruler, but it is from the Lord that a man gets justice.
Proverbs 29: 26
The modern philosopher John Rawls said, "Justice is the first virtue of social institutions, as truth is of systems of thought." I think that Rawls means that justice is fundamental. It is foundational to the formation of our society, and it is an essential platform upon which our society and its systems of governance are formed. Yet it is something that is sadly lacking in our world. Far too often it is held as a tool of power and as a device of oppression. Rulers, governmental and religious leaders, and others in positions of authority and might impose their own concepts of what is right and just upon others in order to bring about conformity and subservient behavior. As a result of this our world is filled with people who live in fear and who, in turn, become fearsome.
Upon consideration of the idea, I think that Rawls is right. Justice is something that should underlie all of the institutions that people use to organize ourselves. It is something that God placed into our world as a part of its creation design. The desire for justice and the capability to grant it are inherent in God's creating humanity in His own image. Justice is a characteristic and a quality of God. I believe that if there were no God, there would be no justice. It is like the other higher qualities of love, peace, mercy, grace, and righteousness. These are qualities that people desire and that benefit us; yet, all of them exist solely because they come from God, Himself. Humanity on its own has a very bad track record in regards to the way that we live together. It is solely through the grace of God that we do find peace, engage in loving others, and seek to govern in a righteous manner. It is that same grace that leads us to justice for ourselves and for others.
If we seek to live in a just world, we must submit ourselves to the King of the Universe. It is essential for us to recognize that we are subjects of an authority that is higher than all other and that is also foundational for all earthly rule and rulers. This reordering of our allegiances is essential for us to become people who treat others justly. Additionally, living in this manner is not learned in government classes or through participation in our processes of governance, it is learned at the foot of the cross of Jesus. Justice is understood as we follow in the steps of Christ while He walked among us and responded to the people and the situations of His world in a manner that brought a living example of God's created desire and intent for just, relational engagement among all people face to face with the real world where we live. In order to know justice, we need to avoid the halls of government until we know the path of Christ. Yet, once we are walking in the steps of the Savior, we should boldly bring Him and His glory into those same halls.


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With the large amounts of gun control legislation being thrown around our nation, people have been writing about prisons, guns, etc… There are several sites that have been keeping an eye on the current state of our fine correctional system, and I would like to take a moment to share those with you.
Basically, as the title so hints, this website is a place where people can post articles about thoughts on the prison system. They have numerous written pieces that highlight the most important events and legislation trending around the correctional systems. Somewhere in here has to be more information about the funding towards our prisons.
Adam Liptak became the New York Times legal reporter in 2002. Previous experience included being Senior Counsel in The Times legal department. He covers the Supreme Court beat, and has provided several articles for The Times regarding the current state of our legal system.
Danny Shaw is the Home Affairs Correspondent for BBC News. He primarily tweets about the current state of our prison system. As you can see from the following tweets, Shaw focuses on the funding and fiscal aspects of our prisons.


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When searching for a new home, you understandably want to find a house that has the ideal amount of space for everyone to feel comfortable. The cost of the home and its style are also top priorities. If your family is like many others, your busy crew is on-the-go regularly. There are some aspects of your new home that can make a life for a busy family more convenient and even more enjoyable. As you search for the perfect house, keep these helpful tips in mind.
Focus on Location
Busy families may have one or two working parents, kids with busy schedules and more. It may seem like you are constantly running to work, heading out to tackle a few errands or shuttling the kids to their activities. When your home is conveniently located close to schools, workplaces, and other frequently-visited venues, you can dramatically reduce the amount of time that you spend in the car.
Continue reading "How to Find the Perfect House That Suits Your Family's Busy Schedule"


text: Over the past year, Eurisy has celebrated its 30 years anniversary. Since its inception, it has relentlessly accomplished its mission and proved to be a highly effective tool for "bridging space and society" through various channels, including the organisation of events. Everyone will agree that the recipe for a successful event is about allowing for networking opportunities or letting discussions sometimes go a little "off-piste". This is the spirit of the series of articles that we initiate today on "Satellites for Sports", aiming at promoting satellite applications for the sports sector in a kind of informal setting.Read more


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CSA/Bioanalyzer: Using the Cellular Labeling and Biomarker instruments of the Bio-Analyzer, the crew initiated an analysis of a control sample.The hardware was powered and the crew ran a control sample. Although the International Space Station (ISS) is equipped with health and life sciences research tools, existing instrumentation for the analysis of biological samples is almost completely lacking as compared to capabilities found in laboratories on Earth. To address this issue, the Bio-Analyzer Commissioning activity tests the Bio-Analyzer facility, which was specifically developed to quantify molecules and cells in body fluids such as blood, urine or saliva. The analysis is conducted on blood samples collected through a finger prick and the instrument provides the data within 2 hours after blood collection.
Combustion Integration Rack (CIR)/Advanced Combustion via Microgravity Experiments (ACME): The crew replaced the ACME Igniter Tip and installed the ACME Short Thermocouple Array, which will allow the continuation of the ACME/Flame Design investigation.The Flame Design investigation studies the production and control of soot to optimize oxygen-enriched combustion and the design of robust, soot-free flames. Soot can adversely affect efficiency, emissions, and equipment lifetime, so this may lead to more efficient and cleaner burner designs. The experiment is conducted with spherical flames of gaseous fuels in the CIR as part of the ACME project. Flame Design is one of five experiments hosted by CIR/ACME.
Cryo chiller Desiccant Remove: The crew removed one desiccant pouch from each of the four openings of the Cryo Chiller Sample Cartridge. Cryo Chiller operates in an EXPRESS Rack and provides a configurable interface at -185°C that accommodates a variety of sample types plus provides flexibility for custom sample configurations if desired. It provides convenient ambient temperature stowage adjacent to the cold volume, so additional hardware may be co-located during crew operations. The unit provides capability for transportation using Commercial Orbital Transportation Services (COTS) vehicles at temperatures currently unavailable (approximately -160°C at 75W).
Fluid Shifts Chibis Ops: Two crewmembers performed the second of two days of their Lower Body Negative Pressure (LBNP) data collection. Fluid Shifts is a NASA investigation, divided into Dilution Measurements, Baseline Imaging, and Imaging with Chibis. The Chibis hardware is used to perform the LBNP part of the experiment. The Fluid Shifts experiment investigates the causes for severe and lasting physical changes to astronauts' eyes. Because the headward fluid shift is a hypothesized contributor to these changes, reversing this fluid shift with a LBNP device is being evaluated as a possible intervention. Results from this study may help to develop preventative measures against lasting changes in vision and prevention of eye damage.
Kidney cells: The crew removed Kidney Cells hardware from Space Automated Bioproduct Laboratory (SABL) and performed the media change inside Life Sciences Glovebox (LSG). Effects of Microgravity on the Structure and Function of Proximal and Distal Tubule Microphysiological System (MPS), or Kidney Cells, examines how kidney health is affected by microgravity and other factors of space travel, including increased chemical exposure, water conservation and recycling, and altered dietary intake. Serious medical conditions caused by poor kidney health, including protein in the urine (proteinuria), osteoporosis, and kidney stones, occur more often and more quickly in space. Knowledge gained can help protect the health of astronauts and also contribute to better treatments for kidney related conditions on Earth.
Mobile Servicing System (MSS) Orbiting Carbon Observatory (OCO)-3: Last night Robotics Ground Controllers powered up the Mobile Servicing System (MSS) and maneuvered the Space Station Remote Manipulator System (SSRMS) and Special Purpose Dexterous Manipulator (SPDM) to extract OCO-3 from the Dragon trunk. The MSS then handed off OCO-3 to the Japanese Experiment Module (JEM) Remote Manipulator System (RMS) which was used to install OCO-3 on the JEM Exposed Facility (JEM-EF). The payload was subsequently activated successfully. Commissioning activities have begun and will continue for 90 days. The OCO-3 will enable carbon dioxide (CO2) observations from the JEM-EF aboard the ISS. The space instrument is designed to investigate questions about the distribution of carbon dioxide on Earth as it relates to growing urban populations and changing patterns of fossil fuel combustion.
Completed Task List Activities:
- None
Ground Activities:
All activities are complete unless otherwise noted.
- OCO-3 robotics
- JEMAL ops
Look Ahead:
Saturday, 05/11 (GMT 131):
- HRF/Repository
- LSG decontamination
- Microalgae
- JAXA/Mouse Mission maintenance
- Crew off duty; housekeeping
- PAO Symbolic
Sunday, 05/12 (GMT 132):
- Astrobee
- Genes in Space-6
- Kidney Cells
- Microalgae
- MVP Cell-01
- Repository
- None
Today's Planned Activities:
All activities are complete unless otherwise noted.
- Kidney Cells Media Kit Retrieve
- FLUID SHIFTS. Connecting Ultrasound Equipment to РБС and РБС power up
- ISS Experience Solid State Drive Changeout
- FLUID SHIFTS. Comm configuration for the experiment
- MICRO ALGAE Culture Bag Daily Deploy
- ISS Experience JEM Setup
- Environmental Health System (EHS) Personal CO2 Monitor Data Transfer and Stow
- On MCC GO FLUID SHIFTS. Gathering and Connecting Equipment for TV coverage
- JEM Airlock Depressurization
- FLUID SHIFTS. Don Chibis
- FLUID SHIFTS. Assistance with LBNP vacuum set (ПВК) CHIBIS and [KMA-01] Medical Hardware Set operations
- Fluid Shifts Ultrasound Service Module Power On And Preparation
- ПХО and DC1 config for EVA
- Fluid Shifts Ultrasound Service Module Scan Operator 1
- Fluid Shifts Ultrasound Service Module Scan Subject
- ISS Experience Introduction Recording
- Kidney Cells Media Change
- FLUID SHIFTS. Doff Chibis
- FLUID SHIFTS. Don Chibis
- FLUID SHIFTS. Assistance with LBNP vacuum set (ПВК) CHIBIS and [KMA-01] Medical Hardware Set operations
- Fluid Shifts Ultrasound Service Module Preparation
- Fluid Shifts Ultrasound Service Module Scan Operator
- Fluid Shifts Ultrasound Service Module Scan Subject
- Environmental Health System (EHS) Total Organic Carbon Analyzer (TOCA) Water Recovery System (WRS) Sample Analysis
- Fluid Shifts RSOS Operations Historical Documentation Photography
- Combustion Integrated Rack Hardware Gather
- FLUID SHIFTS. Doff Chibis
- Bio-Analyzer Sample MELFI Retrieval
- Kidney Cells MELFI Insert 1
- Bio-Analyzer Session Setup Partial
- Kidney Cells Post-Media Change Ops
- FLUID SHIFTS. Deactivation of РБС and Disconnecting USND P/L from РБС
- Health Maintenance System (HMS) ISS Food Intake Tracker (ISS FIT)
- Bio-Anaylzer Maintenance Work Area Preparation
- JEM Airlock Vent
- Bio-Analyzer Microchip Preparation And Control Sample Analysis
- Fluid Shifts Ultrasound Service Module Stow
- Kidney Cells Media Kit Retrieve
- Bio-Analyzer Historical Documentation Photography
- ISS Experience Hardware Relocate
- JEM Airlock Vent Confirmation
- ПХО and DC1 config for EVA
- FLUID SHIFTS. Deactivation of КСПЭ Equipment and Closing Applications on Central Post SSC
- FLUID SHIFTS. Comm reconfig for nominal ops
- Preventive Maintenance of FS1 Laptop (Cleaning and rebooting)
- Public Affairs Office (PAO) High Definition (HD) Config JEM Setup
- Semi-annual BRI maintenance
- PAO Preparation
- Total Organic Carbon Analyzer (TOCA) Sample Data Record
- Combustion Integrated Rack Rack Doors Open
- Public Affairs Office (PAO) Event in High Definition (HD) – JEM
- Combustion Integrated Rack Front End Cap Open
- Food Acceptability Questionnaire
- Kidney Cells Media Change
- ACME Hardware Replace
- Health Maintenance System (HMS) ISS Food Intake Tracker (ISS FIT)
- Testing Inter-modular connectivity (after semi-annual BRI maintenance)
- Emergency RS ISS VHF-1 Comm Checkout from USOS
- Water Resource Management Condensate Tank Offload Initialization
- MPEG-2 Multicast Video Streaming Test, Deactivation of КСПЭ Equipment and Closing Applications on Central Post SSC (without broadcasting vide data to MCC)
- Water Resource Management Condensate Tank Offload Termination
- Fluid Shifts Hardware Final Gather
- Fluid Shifts Ultrasound Hardware Transfer
- Kidney Cells MELFI Insert 1
- Fluid Shifts Hardware USOS Transfer
- Kidney Cells Post-Media Change Ops
- Combustion Integrated Rack Front End Cap Close
- Fluid Shifts HRF PC 3 Reconnect
- Fluid Shifts Ultrasound 2 HRF Rack 1 Setup And Power On From Cargo Transfer Bag
- Combustion Integrated Rack Rack Doors Close
- Combustion Integrated Rack Hardware Return
- INTERACTION-2. Experiment Ops
- Bio-Analyzer Biomarker Microchip Analysis
- Battery Stowage Assembly (BSA) Operation Termination
- CONTROL. Switching Indicator-ISS P/L measuring modes
- Prep for Commemorative Activity
- Metal Oxide (METOX) Regeneration Termination
- MICRO ALGAE Culture Bag Daily Stow
- Cryo Desiccant Remove
- Fluid Shifts Hardware Stow
- ISS Experience Hardware Stow
- Bio-Analyzer Session Conclude


text:
PM of Canada met with Joshua Boyle recently!
Reader comment on item: The Khadrs, Canada's First Family of Terrorism, in the News
Submitted by dhimmi no more (United States), Jan 3, 2018 at 10:51
Dr Pipes
Justin Trudeau met with Joshua Boyle and his family as reported by CTVNews! The report is dated December 19/2017
But this is not the end of the little drama Joshua Boyle has been arrested for 15 criminal charges including sexual assault and uttering death threats and misleading the police
Check this article dated Jan 2/2018
Wow what is the matter with Justin Trudeau? Why would he meet with a sociopath? But again Trudeau said (and I'm paraphrasing) following the Boston Marathon atrocity that we should be understanding and compassionate towards Muslim terrorists and jihadists!
Absolute insanity
Note: Opinions expressed in comments are those of the authors alone and not necessarily those of Daniel Pipes. Original writing only, please. Comments are screened and in some cases edited before posting. Reasoned disagreement is welcome but not comments that are scurrilous, off-topic, commercial, disparaging religions, or otherwise inappropriate. For complete regulations, see the "Guidelines for Reader Comments".
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text:
DescriptionThis mini dress features solid color style, round neckline, cap sleeves, cutout front and back design, sexy mesh splicing front, back zipper closure, and tight stretchy fabric shows off your perfect curves.
Fabric: Blends
Neckline: Round Neck
Sleeve Length: Cap Sleeve
Dress Length: Mini
Embellishment: Cutout
Pattern: Solid Color
Style: Party
Silhouette: Bodycon
Size Chart(Inch) ONE-SIZE Bust 25.2-30.7 Length 28.3 Waist 22-31.5 Size Chart(CM) ONE-SIZE Bust 64-78 Length 72 Waist 56-80
The dimensions data from the measurement of the product, "click here" to see the measuring methods details, and allow 1-2cm (0.4-0.8") differences due to manual measurement, thanks.
Actual fabric colors may vary slightly from online colors due to variations in screen color settings.
30 Days Return + Shipping PolicyShipping & Delivery
Receiving time = Processing time + Shipping time
Note: In orders with more than one item, the processing time will be based on the item with the longest processing time.
We will prepare your order after it have verified. Such as: strict quality control, arrange the suitable carriers.Return Policy
Return within 30 days from the delivered date.
1. Items received within 30 days from the delivered date.
2. Items received unused, undamaged and in original package.


text:
MAC Stylishly Yours Review, Swatches, Photos
I've been mucking around with MAC Stylishly Yours a little bit today. Out of MAC's three upcoming color collections, this is probably the one I was the least excited about even though I tend to love bolder shades of color. With so many color stories coming out at one time I'm leaning towards loving on Peacocky and Cham Pale the most at this point.
How about you?
I only have two pieces from this collection to share but let's have a peek anyway.
MAC Cream Colour Base prove annoying little critters as most shades are not eye safe. Not that you want to run around with some of these shades on your lids but we all like the option to multitask. The one I got was Tickle Me Pink (not eye safe) which is a bright creamy pink. It lends a nice flush of color to cheeks and wears quite well as it has a highly emollient, tacky formula that just stays in place without budging plus it blends easily. Those with dry skin or those living in humid environments will appreciate these.
I also got one of the two Beauty Powder in Play It Proper. This is a soft pink with a bit of subtle sparkle that will work well at brightening up your complexion or even a tiny dab under your eyes to wake them up!
One item that stands apart from everything else in this collection is Dark Diversion Fluidline. MAC describes it as a blackened plum but I get more of a dark bruised cherry. This is stunning and dare I say worth a back up. The shade really brings out my brown eyes and will make not only an incredible eyeliner but also a great base for darker browns, reds, or cranberry and maroon eyeshadows.
MAC Stylishly Yours will have its faithful fans and followers but I think I can safely skip and move on to the other more exciting releases upcoming.
How about you?
Share it!


text: ŠKODA. Servicing with added value. At Capitol ŠKODA we've fixed the prices of all our servicing and maintenance jobs, making it super simple to get superior ŠKODA servicing (try saying that quickly). All this for a budget-busting, easy-peasy price. It's like having your cake and eating it. TO BOOK - Please call us on 01633 730744 Servicing Whether you choose a major service, interim service or an MOT, you can rest assured that the job will be carried out by a factory trained technician who cares about the detail. And when it comes to that all important detail called the price, there's no nasty surprises. TO BOOK - Please call us on 01633 730744 MOT The MOT is an annual test of your ŠKODA's safety, roadworthiness and its exhaust emissions. Available only on UK mainland. All models £39.00 TO BOOK - Please call us on 01633 730744 Oil change service - All models £119 An oil change is carried out every 12 months or 10,000 miles, whichever comes sooner. An oil change service includes: Oil & filter change (fully synthetic oil) Vehicle inspection test and report* Interim service - All models £179 An interim service is carried out every 12 months or 10,000 miles, whichever comes sooner. An interim service includes: Oil & filter change (fully synthetic oil) Vehicle inspection test and report* Full vehicle road test** Vehicle & software enhancements from ŠKODA's database Replacement pollen filter ^ Diagnostic check including report Re-set service interval display Wash and vacuum ŠKODA stamp in service book TO BOOK - Please call us on 01633 730744 Major service - All models £289 A major service is carried out every 24 months or 20,000 miles, whichever comes sooner. A major service includes: Everything from an interim service Replacement air filter ^ Spark plugs ^ (petrol engine) or fuel filter ^ (diesel engine) Remove wheels and check brakes Check/adjust tension of all drive belts Check/replenish gearbox oil (where applicable) Check/replenish final drive oil (where applicable) Check suspension system Check heating/aircon operation TO BOOK - Please call us on 01633 730744 Fixed priced maintenance From a brake pad change to an aircon service, all of our maintenance bookings are carried out by factory trained technicians who know your ŠKODA inside and out. You can think of them like walking-talking ŠKODA encyclopedias. You won't need to read between the lines on your final bill though. The price you see here is the price you'll pay. Simple. AirCon Service - Vehicles built before 30th May 2016^^ £79 AirCon Service - Vehicles built after 30th May 2016^^ £139 AirCon Cleanse £29 Brake Fluid £60 4X4 Transmission Oil £79 DSG Oil Change £189 AdBlue Top-Up £1.50 per litre Battery incl. Stop/Start - Citigo/Fabia £210 Battery incl. Stop/Start - Other models £210 Battery incl. Stop/Start - vRS (Fabia/Octavia) £210 Battery excl. Stop/Start - Citigo/Fabia £120 Battery excl. Stop/Start - Other models £125 Battery excl. Stop/Start - vRS (Fabia/Octavia) £145 Cambelt*** £429 Cambelt and Waterpump £529 Free break check FREE Please note *Excludes Citigo. ^If you are unsure of your vehicle build date, please contact our Customer Service Centre on 0800 333 666 or your local retailer. Prices offered in our Fixed price servicing offer are the maximum selling prices. ŠKODA Authorised Repairers are free to offer prices lower than this offer. Please note - Free Brake Check - With safety-critical components like brakes, it's best to book a check before you start noticing signs of wear. Spongy pedals, squealing or grinding noises, and even your car pulling to one side when the brakes are applied could be signs of brake wear. Book a free brake check with us and our ŠKODA trained technicians will carefully assess all aspects of your system, including brake pads, shoes, calipers, hoses, discs and handbrake linkages. This offer applies to vehicles following a fixed servicing regime only. Customers whose vehicles are on a flexible servicing regime can ask their participating ŠKODA Authorised Repairer about changing over to a fixed servicing regime if they believe this will better suit their needs. For ŠKODA vehicles 3 years (from first MOT) up to 10 years old, excluding V6 models. Fixed price servicing is available to customers at participating ŠKODA retailers only. All prices are (where applicable) inclusive of standard parts, labour, oil and VAT and come with a two-year parts and labour warranty. This offer cannot be used in conjunction with any other offer and may be varied or withdrawn at any time. Further terms and conditions apply, please ask your SKODA retailer for details.


text:
The short answer for this question is that some monoglycerides and diglycerides are vegan, and some are not.
Sorry to say, there's no easy answer that always applies.
The reason is because glycerides are made commercially by a reaction between triglycerides and glycerol.
If you've read my post on if glycerol is vegan, you'll know that some glycerol comes from plant sources, and some comes from animal sources.
To make things worse, the main plant sources are soybeans (clearly vegan) and palm oil (which many vegans avoid).
Takeaway: Most monoglycerides and diglycerides are not vegan, although some are. If you want to be sure, you'll need to contact the company in question, find out where they source their monoglycerides, and then contact that source.
…It's a real headache when you just want to buy a loaf of bread.
Even though glycerides are in a lot of food, you can usually find a similar alternative that doesn't use them. That's my preference.
Where Will You See Mono- and Diglycerides in Food?
Glycerides are in all sorts of things.
But the most common places you'll see them are:
- Bakery products (breads, pies, etc.)
- Icing (e.g. Duncan Hines frosting)
- Soft drinks.
- Ice cream.
- Gum.
- Margarine.
- Candy (e.g. Milk duds)
Bread is the big issue for most vegans. I'd recommend Dave's Killer Bread as a popular bread among vegans if you've having issues finding one.
Most sourdough breads are also vegan.
Verdict: Some Glycerides Are Vegan
Let's summarize the main points from this post.
First, a large portion of monoglycerides and diglycerides are not vegan, but some are.
Some vegans are okay with eating these anyways, as they are only present in very small amounts (typically like 0.1% of the food) – it's your call.
If you want to be sure if a particular food uses vegan glycerides you'll need to contact the manufacturer, get their source, and then ask that source how they make their mono- or diglycerides.
Or, just avoid them, like we do with many other non-vegan ingredients.


text:
Label- Pendleton High Grade Western. Style- Vintage Bomber Aztec Jacket. Condition- Has some minor wear throughout, more noticeable at corners. Also, one pocket lining has been cut on the inside because of a snag. The item "Vtg 70's PENDLETON HIGH GRADE Western Wear Wool Navajo Blanket Jacket Coat L" is in sale since Wednesday, January 2, 2019. This item is in the category "Clothing, Shoes & Accessories\Men's Clothing\Coats & Jackets". The seller is "lstreetboutique" and is located in Griffith, Indiana. This item can be shipped to United States, Canada, United Kingdom, Denmark, Romania, Slovakia, Bulgaria, Czech republic, Finland, Hungary, Latvia, Lithuania, Malta, Estonia, Australia, Greece, Portugal, Cyprus, Slovenia, Japan, China, Sweden, South Korea, Indonesia, Taiwan, Thailand, Belgium, France, Hong Kong, Ireland, Netherlands, Poland, Spain, Italy, Germany, Austria, Bahamas, Israel, New Zealand, Philippines, Singapore, Switzerland, Norway, Saudi arabia, Ukraine, United arab emirates, Qatar, Kuwait, Bahrain, Croatia, Malaysia, Antigua and barbuda, Aruba, Belize, Dominica, Grenada, Saint kitts and nevis, Saint lucia, Montserrat, Turks and caicos islands, Barbados, Bangladesh, Bermuda, Brunei darussalam, Bolivia, Egypt, French guiana, Guernsey, Gibraltar, Guadeloupe, Iceland, Jersey, Jordan, Cambodia, Cayman islands, Liechtenstein, Sri lanka, Luxembourg, Monaco, Macao, Martinique, Maldives, Nicaragua, Oman, Pakistan, Paraguay, Reunion, Uruguay, Russian federation.
- Closure: Zipper
- Brand: Pendleton
- Size Type: Regular
- Size (Men's): L
- Country/Region of Manufacture: United States
- Style: Bomber
- Material: Wool
- Pattern: Navajo
- Color: Multicolor


text: This is the third broadside we produced for New York's annual conference, Typographics 2017. 12" x 17" (304mm x 432mm), printed in black ink on Mohawk Superfine with a typographic key printed on Strathmore Premium Wove duplexed on the reverse side. Featuring (at least) two glyphs per foundry to show the strength of Village as a whole. It will ship Standard Mail in a 17" x 21" mailer to ensure it reaches you without bent corners.


text:
Use annual percentage rate APR, which includes fees and costs, to compare rates across lenders.Rates and APR below may include up to .50 in discount points as an upfront cost to borrowers. Select product to see detail. Use our compare home mortgage loans Calculator for rates customized to your specific home financing need.
Current mortgage rates for August 3, 2019 are still near their historic lows. compare 30-year, 15-year fixed rates, and ARMs to find the best home loan offer all in one place at LendingTree.
Mortgage rates could change daily. actual payments will vary based on your individual situation and current rates. Some products may not be available in all states. Some jumbo products may not be available to first time home buyers. Lending services may not be available in all areas. Some restrictions may apply.
home loans Houston tx We offer a variety of home loan and refinancing options to help ensure your home loan is truly affordable. Your Caliber Loan Consultant can answer your questions and help you apply.. Houston – TX 520 Post Oak Blvd., Suite 455 Houston, texas 77027 nmls# 1151320 state lic# Approved. Meet Our.
Current Mortgage Rates Houston – If you are thinking to refinance your mortgage loan, you can start by submitting simple form online to see how much you can save up. When you know the exact total for your refinance, divide it by the difference between what you currently pay per month and what you will pay per month after refinancing.
Rates Mortgage Property Current Rental For – Current Mortgage Interest Rate For Investment Property At least one more cut is forecast, so now's the time for people to rethink their interest rate. mortgage.
Even more potential buyers may come off the fence: At the current pace of economic and job growth, mortgage rates could hit 6 percent next year. San Antonio, Dallas, Houston and Austin, Texas -.
What should I do today in Houston?
Current rates in Texas are 3.691% for a 30-year fixed, 3.146% for a 15-year fixed, and 3.695% for a 5/1 adjustable-rate mortgage (ARM). Check out our other mortgage and refinance tools Lenders
Other Houston mortgage types mixed. The average rate for 15-year fixed-rate mortgages rose 2 basis points, growing to 3.55 percent. Monthly payments on a hypothetical $165,000, 15-year fixed-rate mortgage ticked up along with rates, scaling up $1.62 to $1,183.61.
Mortgage delinquency rates continued to fall in October, with the national rate ticking down 0.1 percent and the percentage of non-current loans declining in. Texas markets such as Houston,