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Technologies to Enable Autonomous Detection for BioWatch: Ensuring Timely and Accurate Information for Public Health Officials: Workshop Summary (2014)
Chapter: Appendix H: White Paper 3: State of the Art for Autonomous Detection Systems Using Immunoassays and Protein Signatures
« Previous: Appendix G: White Paper 2: Nucleic-Acid Signatures at Three Levels of Readiness for BioWatch
Suggested Citation:"Appendix H: White Paper 3: State of the Art for Autonomous Detection Systems Using Immunoassays and Protein Signatures." Institute of Medicine and National Research Council. 2014. Technologies to Enable Autonomous Detection for BioWatch: Ensuring Timely and Accurate Information for Public Health Officials: Workshop Summary. Washington, DC: The National Academies Press. doi: 10.17226/18495.
State of the Art for Autonomous Detection
Systems Using Immunoassays and Protein
R. Paul Schaudies, Ph.D.
GenArraytion, Inc.
A white paper prepared for the June 25–26, 2013, workshop on Strategies for Cost-Effective and Flexible Biodetection Systems That Ensure Timely and Accurate Information for Public Health Officials, hosted by the Institute of Medicine's Board on Health Sciences Policy and the National Research Council's Board on Life Sciences. The author is responsible for the content of this article, which does not necessarily represent the views of the Institute of Medicine or the National Research Council.
Antibody-based strategies dominated the biological agent detection landscape during the 1980s and well into the 1990s. Antibodies have been used by the U.S. military for the detection of biological warfare agents since the first Gulf War. The ability of immune systems to identify and synthesize immunoglobulins against non-self-structural elements, primarily proteins, with excellent sensitivity and specificity is the backbone of antibody-based detection and identification approaches.
The introduction of nucleic acid–based detection and identification technologies, primarily nucleic-acid amplification technologies, in the later 1980s presented an ability to look at the genetic information in addition to the structural elements of pathogenic organisms. While systems such as BioWatch have focused almost exclusively on nucleic-acid amplification technologies, the continued development of antibody- and
protein-based recognition systems warrants a fresh look at the state of maturity of structural recognition approaches to biological identification. This paper highlights some of the more mature and promising technologies that could be adapted by the BioWatch Program for the identification of harmful biological agents.
All of the systems discussed in this paper can meet or exceed the detection requirements established for the next generation of BioWatch. The advantage of antibody-based and other protein-based technologies is speed. Many of these systems provide answers in 3 to 10 minutes and can operate in a continuous fashion at reasonably low cost with minimal infrastructure support. Some of the systems can reach sensitivities approaching that of nucleic-acid amplification.
The Department of Homeland Security (DHS) Office of Health Affairs is researching the potential to employ a fully autonomous networked biodetection capability that will be deployed, operated, and sustained, both indoor and outdoor, in selected BioWatch jurisdictions throughout the United States to continuously monitor the air for agents of biological concern. This requirement supports the DHS plans to increase the capability of the BioWatch system by augmenting and ultimately replacing the current collection and biodetection capability with an autonomous biodetection capability that will improve timeliness, time resolution, population coverage, and cost-effectiveness while enabling the program to stay within its fiscal constraints.
The autonomous detection capability must be able to (1) rapidly process and accurately analyze aerosol samples with a high level of confidence; (2) automate and integrate the major system functions into the detector, including aerosol sample collection, preparation, analysis, and analytical results reporting; (3) operate in its intended indoor and outdoor environments; and (4) disseminate and archive analysis results and system operational data via the C3 network, known as the BioWatch Gen-3 Operations Support Service. The requirement is for autonomous biodetectors, for both indoor and outdoor use, that continuously monitor the air for agents of biological concern (24 hours per day, 365 days per year) and all necessary information technology, equipment, consumables, and technical support for the nationwide deployment, operations, and maintenance of this autonomous biodetection capability.
Appropriateness to BioWatch Problem
The use of antibodies, or immunoglobins, as a structural recognition element in a biological detection architecture takes advantage of one of the body's premier defense systems. Antibodies represent the body's ability to discriminate between self and non-self. The standard IgG immunoglobin is a Y-shaped molecule with the short arms representing identical binding elements and a base that can be anchored to a variety of surfaces. The binding region recognizes a small region, generally three to five amino acids in size, which is unique from the host that generated the antibody as part of an immune response. A single immune cell makes a single antibody structure generating "monoclonal" antibodies. The collection of individual antibodies from blood of a host represents a multitude of these individual antibodies and is termed polyclonal, each individual antibody targeting a unique structure.
The antibodies are responsible for binding foreign entities to target them for destruction and removal from the body. The binding is specific and sensitive. Normal ranges of disassociation constants (indicating tightness of fit) range from 10–9 to 10–11. These values are important because they are central in determining the sensitivity and specificity of a detection system utilizing the antibody as a detection element.
Biological identification systems rely on the collection and detection of recognizable entities that are unique to the harmful biological agent and not found independent of it. The BioWatch Program is designed to protect against a catastrophic event caused by bacteria or viruses. Toxins are also a concern because they are potent at very low levels and represent a challenge to nucleic acid–based detection systems. The argument that nucleic acids may contaminate the preparation has merit, but antibody-based systems are more effective and consistent than nucleic acid–based systems for the detection and identification of toxins. In addition, there are some antibody-based systems that provide detection and discrimination of microorganisms at levels that support the autonomous detection requirements established for BioWatch Gen-3.
Sample Collection and Processing
Sample collection and processing are the ultimate driver for sensitivity. The fundamental objective for biological sample collection and preparation is to extract the target signatures from the environment into the volume required for one or more specific assays. A front-end air-
sampling device typically removes particles of defined size and mass into a collection matrix, generally an aqueous liquid. This liquid undergoes extraction(s) of various types to separate the biological signature(s) of interest from the clutter in the sample. As tests are run on smaller volumes, the concentration of target must increase, given a fixed level of detection. If a given test has a sensitivity of 100 signatures, then the concentration (signatures/ml) is indicated in Table H-1. The signature may be a genetic sequence, an epitope for antibody binding, or a mass spectrometer target.
If the copy number required for detection decreases by a factor of 10, then all of the concentration numbers also decrease by that factor. This simple table highlights the requirement for efficient collection and processing for analysis of biological targets.
Signature copy number from a single organism can be larger than the actual copy number of the organism in multiple ways. Most of the antibodies are directed against antigenic determinants that are present on the surface of the cell. Generally there are hundreds to thousands of copies of these structures on a single bacterium or virus. If the target organisms are sonicated to release the soluble contents, hundreds to thousands of membrane fragments can be released from a single organism (each behaving as a separate entity), providing many more independent binding opportunities and dramatically increasing the sensitivity of the system. In this way antibody detection and identification can compete with sensitivities achieved by nucleic-acid amplification technologies.
TABLE H-1 Required Concentrations
Copy Number Required
Copy Number/ml Required
1.0 ul
1.0 pl
1.0 fl
NOTE: fl = femtoliter; ml = milliliter; pl = picoliter; ul = microliter.
Antibody–Antigen Interaction
The vast majority of detection technologies that employ antibodies as structural recognition elements involve initial immobilization of a capture antibody on a solid substrate. This substrate can be a membrane, as in traditional lateral flow assays. Alternatively, an optically transparentmatrix can be used for evanescent wave utilization of suspended substrates, such as a bead used in magnetic separations or optical laser discrimination.
Figure H-1 illustrates a basic approach, referred to as a sandwich assay, for the utilization of antibodies. The primary, or capture, antibody binds the target in such a way that the target is then associated with the capture substrate.
Both antigen and reporter antibody binding events are critical to the fidelity of the detection system. Nonspecific binding is caused by a label, or reporter molecule, being localized with the capture antibody, and it creates noise in the system. It is generally accepted that a specific signal must be at least three times the intensity of the background to be a specific binding event.
FIGURE H-1 Visual representation of the sandwich-style assay in which the capture antibody (pink) is immobilized to a substrate. The red HMGB1 represents the target that binds the capture antibody and is subsequently bound by the blue reporter antibody.
NOTE: ELISA = enzyme-linked immunosorbent assay; HMGB1 = high-mobility group box 1 protein.
SOURCE: http://dc184.4shared.com/doc/goS0-D6B/preview.html.
The reporter molecule is a readable signal that represents what is measured by the system and is intended to be a function of amount of target present in the sample. A wide variety of reporter systems are used in antibody-binding detection methods and represent a significant variable in the sensitivity of various systems. The types of reporter systems can be divided into three broad categories. The first is optical reflectance, such as the colloidal label on secondary antibodies utilized in most lateral flow devices. The second is fluorescence whereby a reporter molecule is excited at one wavelength and then emits light at a different wavelength. The spectral and temporal excitation and emission differences allow various methods of collection to enhance the signal-to-noise difference. The third method is an enzymatic reaction whereby a nonvisible substrate is converted into a detectable product. The enzymatic products vary with different systems.
This section presents a variety of different platforms, all of which have been incorporated into a product that can reach TRL 6-plus between 2016 and 2020.
The Luminex platform can accommodate both antibody-based and nucleic acid–based biological detection schemes. It is presented first because it is relatively mature and is incorporated into the current BioWatch Program. The Gen-2 BioWatch Program has used the Luminex LX-200 platform to run a polymerase chain reaction (PCR) nucleic-acid amplification–based test. The LX-200 is also integrated in an autonomous PCR-based Gen-3 prototype that has undergone substantial field testing. A modification of the Gen-3 prototype includes the newer MagPix instrument that employs magnetic beads and in which a robust charge-coupled device (CCD) camera replaces the flow cytometer in the LX-200. For antibody-based detection, Luminex uses a bead-based platform in which the capture antibodies are covalently linked to color-coated beads. The BioWatch Program uses the LX-200 system, which incorporates a laser and a complex fluidics system. For future systems
the newer MagPix instrument is recommended as it eliminates the flow cytometer laser and complex fluidics. Figure H-2 illustrates the Luminex open architecture platform that allows for the incorporation of new assays into an existing menu of tests.
Figure H-3 illustrates the detection mechanism for the MagPix system, which uses LEDs for bead optical interrogation and immobilizes the beads with a magnet to allow for increased image capture time, thereby improving sensitivity.
Figure H-4 shows the Magpix system, which has a relatively small footprint and has user-friendly interfaces.
The MagPix uses a ruggedized hardware platform that does not require a company-trained individual to conduct operations. Manufacturer provider performance is provided in Table H-2.
FIGURE H-2 Illustration of the multiplexing capability of existing Luminex antibody-based assays for biothreat targets.
FIGURE H-3 Illustration of the optical imaging system of MagPix.
NOTE: CCD = charged-coupled device; LED = light-emitting diode.
FIGURE H-4 Image of the complete MagPix system with computer interface.
TABLE H-2 MagPix Performance
Multiplexing capability
Up to 50 individual analytes
Reagent compatibility
MagPlex® magnetic microspheres
3.5 logs
Microplate type
96-well plate
96 wells ≤60 mins
Sample temperature control
35°–60°C (95°–131°F)
Sample volume uptake
20–200 ml
Probe piercing
Auto adjust-probe height
Daily startup
≤15 minutes
Approximately 106 copies of DNA or single-digit picogram levels of protein, 102–103 copies of bacteria.
The open architecture allows for significant flexibility for generation of additional tests in any laboratory. Significant field experience and design engineering experience.
Only a fraction of the beads incubated with target are analyzed, which dilutes the signal.
PathSensors, Inc.
PathSensors, Inc. (PSI) manufactures reagents and instrumentation for the detection of pathogens in indoor and outdoor environments, with a focus on biodefense and agri-food applications. Detection of pathogens is based on the CANARY® technology originally developed at Massachusetts Institute of Technology's Lincoln Laboratory (Science, 2003) and licensed exclusively to PSI for these applications.
The CANARY® technology shown in Figure H-5 differs significantly from the standard immobilized antibody. This is the only technology discussed in this paper that exploits the natural mechanism of antibody response when a specific antigen is detected. The antibodies are expressed in B lymphocytes that are the natural display mechanism within the host. Upon formation of a dimer with the antigen serving as the bridge, the antibodies stimulate a cascade of intracellular events. In this process, the cascade is linked to the generation of photons, which are detected using a photomultiplier tube. The process forces antigen and antibody together by centrifugal force and the response is as specific as the antibody and very rapid. The complete response occurs in less than 5 minutes.
FIGURE H-5 Principle of the CANARY® Biosensor. B-cell lines are engineered to express membrane-bound antibodies that trigger the release of intracellular calcium upon specific interaction with their target. These intracellular signaling processes rapidly culminate in the activation of bioluminescent proteins to generate photons of light within seconds after contact between the biosensor and its cognate pathogen.
SOURCE: Pathfinder CEO.
The principal benefits of the CANARY® technology include the speed of detection, which takes as little as 2 minutes, and the sensitivity that is conferred by the intracellular signal amplification, which allows detection of very low levels of target. In analytical assays, CANARY® demonstrates the ability to detect as few as 10–100 colony-forming units (cfu) of target organisms with a time to result of 3 minutes, including sample processing (see Figure H-6).
This technology has been fielded in one or more Department of Defense buildings for over 5 years with excellent results. The monitoring system operates with a trigger from a particle detector. The system has had thousands of collections and tests without any false positives over the 5-year period of operation. The system is fully capable of adding additional targets by the incorporation of different antibody genes into the progenitor B-cell line. This technology is a definite candidate for BioWatch, either as a trigger or as an integrated detection/identification system.
Strengths: A significant strength of this approach is speed, with results within minutes with good sensitivity.
Weakness: Reagent stability is a concern because the B cells must be viable in order to respond. Anything in the collected sample that harms the integrity of the B cell could cause a problem.
FIGURE H-6 Sensitivity and speed of the CANARY® technology. In analytical assays, CANARY® enables detection of 10 cfu of B. Anthracis (Sterne) spores in only 3 minutes.
NOTE: RLU = relative luminescence unit.
TABLE H-3 Selected Subset of Available B-Cell Lines with Analytical Sensitivities
Agent Antigenic Target Test Panel Assay Sensitivity
Bacillus anthracis (spores) Targets anthrax spore coat antigen Positive against live Sterne strain, gamma-killed and live Ames strain 10 cfu/sample (Sterne) <100 cfu/sample (Ames)
Yersinia pestis Targets F1 antigen Tested against inactivated Yp CO-92, and live Kim5 strains 50 cfu/sample
Francisella tularensis Targets cell wall polysaccharide Positive for inactvated Ft Schu4, LVS <100 cfu/sample
Smallpox Orthopox specific Tested against live vaccinia virus, inactivated vaccinia virus (Lister strain) <100 pfu/sample (UV-Psoralen inactivated virus) 500 pfu/sample (live virus)
Brucella spp. Tested against B. suis, B. abortus, B. melitensis 50 cfu/sample
Venezuelan equine encephalitis (VEE) virus Targets VEE virus capsid protein Positive against killed VEE3880, Menall, P676, TC-83, PTF-39, and Trinidad donkey 500,000 pfu/sample Strain variability in LoD
Ricin Ricin A chain Tested against active ricin <0.4 ng/sample
Botulinum toxin BoNT/AHc Tested against active botulinum toxin A 16 pg/sample against active toxin
Bacillus subtilis spores Tested against live spores 50 cfu/sample
NOTE: BoNT/AHC = heavy chain of botulinum neurotoxin serotype A; cfu = colony-forming unit; LoD = limit of detection; LVS = live vaccine strain; ng = nanogram; pfu = plaque-forming unit; pg = picogram; spp. = species; UV = ultraviolet.
Research International, Inc. (RI) has been involved in the BioWatch program for a number of years. It supplied a portion of the air sampling system to Lawrence Livermore National Laboratory during development of the autonomous pathogen detection system. Since then it has been a subcontractor to Northrop Grumman on the Gen-2 program and has manufactured the entire aerosol collection system for all the units delivered to date.
The primary operating costs for a BioWatch Program accrue from the biological assays needed to detect and identify pathogens. The assay chemistries required and the labor involved in frequent servicing are expensive. To minimize these costs one could implement a strategy of monitoring bioaerosol concentrations with an optical biofluorescence instrument that has no consumables and minimal maintenance. Only when this instrument detects a rise in aerosol concentration would collection and wet assay identification be initiated.
The TacBioHawk is an integration of the TacBio biological particle detector and the BioHawk aerosol collector and bioidentifier into a single package. It is designed for continuous outdoor monitoring of biothreat agents over a wide range of environmental conditions.
The biological particle detector uses ultraviolet-excited fluorescence to monitor biological particle levels in the air and elastic scattering to monitor other types of particles. This particle detection portion of the TacBioHawk is extremely useful for tracking background levels of airborne materials and providing an alarm if there is an unusual increase in biological aerosol levels. However, it does not identify the type of biological material detected, and for that reason it is correctly characterized as a "trigger."
The BioHawk portion of the system provides identification in the event of a TacBio alarm. When the BioHawk is triggered into operation by the TacBio, an aerosol sample is automatically collected and an eight-agent sandwich format fluoroimmunoassay is performed. The entire process can be completed with no human intervention within a 20- to 30-minute period. Additional samples can be collected and assayed within 17 minutes.
TacBio Biodetector Module
The TacBio aerosol biodetector's mode of operation is based on the well-known fact that non-biological materials primarily absorb and scatter impinging ultraviolet (UV) light, whereas biological materials fluoresce as well as scatter UV light. With the recent development of GaN semiconductor light-emitting diodes, high-brightness UV sources have become available that are energy efficient and more reliable and temperature tolerant than current UV lasers (<400-nm wavelength). In addition, very-high-speed analog electronics are used to count the individual photons emitted by each particle, providing a "Geiger counter" operating mode that eliminates most of the drift and aging problems seen with analog optoelectronics. The TacBio requires little maintenance and no consumables, and it is an ideal first line of defense for monitoring areas that cannot be accessed regularly.
Although aerosol data can be transferred to a remote location for analysis and storage using the TacBioHawk's serial RS-232 data line, it is also stored internally on a removable SD memory card within the unit. This card can store up to 5 years of aerosol alarm data.
BioHawk Module
The BioHawk® is a portable eight-channel bioassay system with automated sample collection capability that is suitable for high-sensitivity monitoring of live agents and protein toxins. Once activated by the TacBio, the BioHawk® collects aerosol particulates from the surrounding air with its wetted wall cyclone air sampler and automatically transfers the concentrated sample from the air sampler to a bioassay module. This module performs automated sandwich-style antibody assays in eight separate detection channels over a period of 15 to 20 minutes, after which results are communicated to users through a touch-screen liquid-crystal display and by wireless or RS-232 link to personnel at a remote location. System operation may also be remotely controlled in real time with virtually every fluid or air handling function being capable of remote manipulation.
The bioidentification portion of the instrument can also analyze fluids from other sample collection systems or from liquid samples loaded into the instrument through a manual sample port. A built-in microcomputer provides unparalleled operating flexibility and sophistication.
Bioassays are performed within a disposable, credit card–sized, plastic assay coupon. Unlike other technologies on the market that are based on a "use once and discard" philosophy, this coupon and the reagents stored within it may be used for 10 assay procedures before being discarded. Because the coupon can execute up to 8 different assays simultaneously, a total of 80 individual assays may be performed before the coupon is discarded. This capability can substantially reduce life-cycle cost.
While possessing a high level of function and great versatility, the unit is still very easy to use. Most global functions such as air sampling and bioidentification are performed using multistep protocols (recipes) developed by RI and stored in the system's computer memory. The unsophisticated user needs only the most fundamental level of training since the internal processes and steps are preset through the built-in computerized recipes. For more sophisticated users, bundled Windows-based software allows the development of customized sample collection and detection protocols.
The system is designed to be operated over a very wide temperature range, from –32°C to 60°C. In low-temperature environments sampled air is preheated to above freezing before entering the wetted wall cyclone sampler. At high ambient temperatures, cooling fans circulate air into the transport case to prevent overheating. Bioassay reagents are maintained in a very stable lyophilized (freeze-dried) form until they are needed and are only rehydrated when a bioassay is to be performed, greatly extending the assay coupon's useful life. Once hydrated, reagents are useable for a period of up to 48 hours.
The current version of the TacBioHawk was developed for the armed forces of a NATO country, which specified using only a limited amount of power for the unit. Because air temperatures below freezing require heating the incoming air for operation of the wetted wall cyclone and for operation of the immunoassays, the air sampler was limited to 40 liters per minute (lpm). However, access to mains power will allow operation with a 325-lpm aerosol collector. If this volume proves insufficient, RI's SASS 4000 aerosol preconcentrator, which operates at 3600 lpm, can be integrated into the system. It would increase overall sensitivity by roughly a factor of 10.
The characteristics of the TacBioHawk are as follows:
• Sensitivity
o 8 × 104 cfu-min/m3 for Bacillus anthracis spores with 325 lpm aerosol collection.
o Probability of detection of 95 percent for subspecies dependent on the particular Department of Defense Critical Reagents Program antibodies used.
• Specificity
o False-positive rate 1 to 2 percent for 17-minute assay with a limit of detection of 5 × 104 cfu/mL for B. anthracis.
o Speciation is dependent on quality of C-reactive protein (CRP) antibodies
• Flexibility
o Current hardware supports simultaneous assays for one to eight agents in the same 2-mL sample. Addition of a second detection module would increase range to 16 agents. A maximum of 20 agents is possible.
o Quantitative information is saved in on-board flash memory and may be automatically transmitted to a remote computer if desired.
• Measurement interval
o Aerosol trigger: 1 minute updates
o Sample collection and agent identification initiated on alarm from aerosol trigger
o Sample collection: 5 minutes minimum recommended for first sample after alarm
o Agent identification: 30 minutes assay time for first sample after first collection
o Sample collection: user definable after initial collection
o Agent identification: 17 minutes for each additional sample
• Cost
o Initial purchase: $125,000 to $150,000 in quantities <10 depending on exact configuration. Significant discounts available for larger volumes.
o Operational maintenance: <$1,000/year hardware; ~3 hours/quarter staff time for cleaning.
o Consumables: $250 per assay coupon. Coupon only used if assay is triggered. Once triggered, up to 10 assays may be run.
o Once set up on site, the TacBioHawk is fully automated. Only maintenance and replacement of consumables required.
o Mean time between failures: >10,000 hours with regular maintenance.
o Availability: currently 99.9 percent or better depending on environment.
• Operational environments
o Can operate in dirty internal environments
o Current design can operate –32°C to 60°C. Operation between 50°C and 60°C is permitted up to a total time of 1,000 hours.
Strengths: This system has extensive field testing.
Weaknesses: Specificity of antibodies may not provide needed discrimination.
Battelle REBS Biosensor Technology
Battelle has patented an integrated collection and identification spectroscopic technology, the Resource Effective Bio-Identification System (REBS), which provides a portable platform that is a lightweight, low-cost, networked, battery-operable system with near-real-time identification of environmental aerosols, surface contamination, and waterborne biological warfare agents (BWAs) (see Figure H-7). The process uses Raman spectroscopy for BWA identification and has undergone extensive government field testing over several years. By using a spectroscopic method, REBS significantly reduces the amount of consumables and overall life-cycle cost, reduces the amount of time to identify agents, and includes an agile software-updatable identification capability for emerging threats. REBS autonomously processes samples from collection to detection/collection subsystems through to identification, thus removing errors that are often encountered in manual sample processing methods. REBS delivers an archive confirmatory sample via a removable vial compatible with genetic and other confirmatory methods. REBS delivers performance for approximately $0.05 per sample. This drastic reduction in identification cost versus that of traditional systems allows for contin-
uous collection and identification of BWAs while eliminating the burden of reagent usage.
Probability of Correct Identification
REBS demonstrated a probability of correct identification above 97.5 percent for the samples supplied by a government sponsor for an identifier technology readiness evaluation. During limit-of-identification testing at Battelle Memorial Institute, all of the aerosol challenge samples were identified correctly. Based on the number of trials in this test, the 95 percent confidence bounds on the probability of correct identification are 96.5–100 percent. These tests included four BWAs, including gram-negative and gram-positive bacteria, virus, and protein simulants.
Battelle has carried out extensive testing to establish the probability of correct identification, first based on Raman spectroscopy of single particles and, second, on the consensus of several identified particles
FIGURE H-7 Battelle's Resource Effective Bio-Identification System (REBS) provides collector, detector, and identifier functions within a single, portable system.
SOURCE: Provided by Battelle.
within an aerosol collection. The probability of correct identification for a single particle has consistently performed to greater than 90 percent for any agent within an extensive spectral database. However, a system alarm utilizes several particles to form a consensus on the identity of a collected aerosol sample. Based on the consensus result, the system had been configured to identify at least three particles to confirm the aerosol identity. By accumulating information over these three particles, the probability of correct identification for a sample is above 97.5 percent for all agents tested.
Detection and Identification False-Positive Rates
REBS has been extensively tested concerning its detection and identification false-positive rates in realistic environments. The REBS false-positive rate has been measured over several years in both indoor and outdoor locations throughout the United States. In government-sponsored tests, REBS demonstrated 1,629 hours of outdoor operation without a reported false-positive, at Ft. Bliss in El Paso, Texas, during July 2009. To date these tests correspond to 9,774 distinct samples with a 10-minute analysis interval. More recently, three REBS systems were tested in the Boston subway system from October 2012 through April 2013. In this test REBS demonstrated 2,454 hours of operation in the subway system without a reported false-positive. These tests correspond to 5,860 distinct samples with a 15-minute analysis interval.
No samples in any trial have been identified as one of the targets within the system database, and therefore the probability of false-positives currently has an average of 0.0 percent with an upper bound of 0.001 percent (80 percent confidence interval). Naturally occurring biological particulates are routinely detected in these tests, but none has been misidentified as Bacillus anthracis, Francisella tularensis, Venezuelan equine encephalitis, Clostridium botulinum toxin, or any other material within the system database.
REBS is 1.5 cubic feet and weighs 38 pounds. Its dimensions are 18 × 12 × 12 inches. REBS requires electrical power from at least one source. Potential power sources include military-style batteries, 120- to 240-VAC (volts alternating current) at 50- to 60-Hz (hertz) facility power, or 21- to 32-VDC (vehicle dynamics control) vehicle power.
System Communications
REBS is locally controlled via a simple push-button keypad. Alarms, warnings, battery life, system status, configuration, and meteorological conditions are viewable by a liquid crystal display interface adjacent to the keypad. Remote control and monitoring are achieved using a wired or wireless network communication protocol and remote computer graphical user interface.
Expandable Threat Identification
REBS detects and identifies at the genus, species, and strain levels based on the Raman signatures generated by a laser field interacting with the molecules and structural conformations of biological materials that comprise BWAs. Battelle currently maintains more than 120 spectral signatures in an on-board database, which is easily and cost-effectively expandable to include new threat materials through a simple software patch update via wireless connectivity. As a result, hardware modifications, system recalibrations, perishable assays, and costly reagents are not required to expand the functionality of REBS. Battelle has constructed a library of signatures for various spores, bacteria, viruses, toxins, common battlefield interferants, chemical warfare aerosols, and some explosives. Expanding threat identification signatures takes as little as 24 hours and has been tested in government-sponsored technology readiness evaluations. Once the signature is determined, it can be sent wirelessly to fielded systems, enabling instantaneous enhanced threat warning capabilities.
Analysis Interval
The limit of identification for the REBS system is dependent on the analysis time interval. An increase in the time available for analysis increases the sensitivity of the system. An analysis time of 20 minutes is approximately two times more sensitive than the same system configured for a 10-minute analysis time. The additional analysis time allows for more materials to be collected and more of the sample to be analyzed. The ability to change the REBS duty cycle represents a unique capability; the duty cycle can be adjusted via software configuration changes. This attribute can be desirable for the various situations where the threat likelihood may be higher than normal. In government-sponsored testing, REBS was configured to collect and analyze samples with a 20-minute
duty cycle. In this test REBS demonstrated a 23-minute time to result at a BWA simulant concentration of approximately 200 particles per liter of air.
System Cost and Consumables
REBS is a reagentless BWA identification technology that eliminates the need for expensive and perishable reagents and consumables. REBS is significantly less expensive to operate than any other equivalent technology, based on logistical burden expense reduction alone. Moreover, the REBS ability to add new threat agents via software offers a compelling value over similar technologies that are assay- or wet chemistry–based.
Strengths: Cost per analysis is low, rapid cycle time, nondestructive analysis of sample.
Weakness: Customers are not familiar with technology, therefore a significant barrier to entry.
MesoScale
The MesoScale technology is a mature system with many systems in operation. The electrochemiluminescence system offers significant sensitivity and was once a system of choice for the U.S. Army. The system provider was purchased, and the older systems are no longer fully supported by the new owner. Antibody-based assays immobilize the capture antibody in a black plastic well, and then standard sandwich assays are performed with a proprietary reporter light-generating reaction.
Electrochemiluminescence detection uses labels that emit light when electrochemically stimulated (see Figure H-8). Background signals are minimal because the stimulation mechanism (electricity) is decoupled from the signal (light).
Labels are stable and nonradioactive and offer a choice of convenient coupling chemistries. They emit light at ~620 nm (nanometer), eliminating problems with color quenching. Few compounds interfere with electrochemiluminescent labels, so one can use large, diverse libraries with confidence. Multiple excitation cycles of each label amplify the signal to enhance light levels and improve sensitivity.
FIGURE H-8 Representation of the electrochemical light-generating reporter system for MesoScale technology.
NOTE: MSD = Meso Scale Discovery; Ru(bpy) = ruthenium tris difulleride; TPA = tripropylamine
SOURCE: Company website.
FIGURE H-9 Multispot plates, which offer arrays within the well for increased throughput and assay multiplexing, are available in 24-, 96-, and 384-well formats, with up to 100 spots per well.
The system offers multiplexed detection technology by spatially separated spots on the bottom of a 24-, 96-, or 384-well format microtiter plate (see Figure H-9).
Sensitivities of the MesoScale systems are generally superior to standard ELISA assays. The sensitivities achieved by currently available systems are adequate for the BioWatch requirements.
Strengths: Sensitive reproducible tests. Proven technology.
Weakness: Not currently configured for BioWatch operations.
TECHNOLOGY FOR FUTURE APPLICATIONS
Quanterix Corporation Simoa Technology
Quanterix has developed, and is commercializing, a novel antibody-based technology that makes some fundamental changes when compared with existing approaches. The figure below provides a cartoon diagram of the various steps from sample collection to readout of results.
Antibodies are covalently attached to microspheres and then incubated with sample in such a way that the ratio of bead to target is slightly less than unity. The goal is to have only one target per bead. All reactions are in solution phase, which speeds the binding reaction compared to antibodies fixed on a planar surface. The beads occupied with target are then incubated with a secondary reporter antibody, which is covalently linked to an enzyme.
The conjugated enzyme bead complex is rinsed in a solution containing an optically inert substrate. The bead–substrate mixture is floated across a plane of microwells, each of which can hold only one bead. The wells are then sealed and the enzyme converts substrate into fluorescent product. Each fluorescent well represents a single target molecule. The process is diagramed in Figure H-10. The result is a quantified number of targets with sensitivities up to over 1,000-fold demonstrated with existing technology.
The current instrument, pictured in Figure H-11, is a floor standing unit.
While the biodefense industry may not be a current market for Quanterix, the technical approach offers some significant improvements over existing technologies. This type of approach should be considered for post-2020 fielding.
FIGURE H-10 The Quanterix antibody-based technology. (a) Single protein molecules are captured and labeled on beads using standard ELISA reagents. (b) Tens of thousands of beads—with or without immunoconjugate—are mixed with enzyme substrate and loaded into individual femtoliter-sized wells. The microwells are sealed with oil. (c) Fluorophore concentration in the small sample volume of wells containing the target analyte rapidly reach detectable limits using conventional fluorescence imaging and can be digitally counted. The percentage of beads containing labeled immunocomplexes can be computed at low concentration because they follow a Poisson distribution; at higher concentrations the intensity of the aggregate signal provides an analog measurement.
FIGURE H-11 Quanterix Simoa HD-1 analyzer.
All of the systems presented have the capability to meet the majority of the autonomous detection system requirements. The most significant challenge will be the ability to provide sufficient specificity for organism discrimination. A clear example of this is that a Food and Drug Administration–approved antibody-based test for Bacillus anthracis, sold by Tetracore, Inc., uses the harmless vaccine strain for a positive control. This challenge may be overcome by the speed in which these technologies provide information to the customer community.
Next: Appendix I: White Paper 4: State of the Art for Autonomous Detection Systems Using Genomic Sequencing »
Technologies to Enable Autonomous Detection for BioWatch: Ensuring Timely and Accurate Information for Public Health Officials: Workshop Summary Get This Book
The BioWatch program, funded and overseen by the Department of Homeland Security (DHS), has three main elements--sampling, analysis, and response--each coordinated by different agencies. The Environmental Protection Agency maintains the sampling component, the sensors that collect airborne particles. The Centers for Disease Control and Prevention coordinates analysis and laboratory testing of the samples, though testing is actually carried out in state and local public health laboratories. Local jurisdictions are responsible for the public health response to positive findings. The Federal Bureau of Investigation is designated as the lead agency for the law enforcement response if a bioterrorism event is detected. In 2003 DHS deployed the first generation of BioWatch air samplers. The current version of this technology, referred to as Generation 2.0, requires daily manual collection and testing of air filters from each monitor. DHS has also considered newer automated technologies (Generation 2.5 and Generation 3.0) which have the potential to produce results more quickly, at a lower cost, and for a greater number of threat agents.
Technologies to Enable Autonomous Detection for BioWatch is the summary of a workshop hosted jointly by the Institute of Medicine and the National Research Council in June 2013 to explore alternative cost-effective systems that would meet the requirements for a BioWatch Generation 3.0 autonomous detection system, or autonomous detector, for aerosolized agents . The workshop discussions and presentations focused on examination of the use of four classes of technologies--nucleic acid signatures, protein signatures, genomic sequencing, and mass spectrometry--that could reach Technology Readiness Level (TRL) 6-plus in which the technology has been validated and is ready to be tested in a relevant environment over three different tiers of temporal timeframes: those technologies that could be TRL 6-plus ready as part of an integrated system by 2016, those that are likely to be ready in the period 2016 to 2020, and those are not likely to be ready until after 2020. Technologies to Enable Autonomous Detection for BioWatch discusses the history of the BioWatch program, the role of public health officials and laboratorians in the interpretation of BioWatch data and the information that is needed from a system for effective decision making, and the current state of the art of four families of technology for the BioWatch program. This report explores how the technologies discussed might be strategically combined or deployed to optimize their contributions to an effective environmental detection capability.
1 Introduction 1–6
2 Overview of the BioWatch Program 7–22
3 Public Health's Perspective on the Role of BioWatch in the Decision-Making Process 23–40
4 Potential Technologies for the BioWatch Program 41–80
5 Final Thoughts 81–94
Appendix A: References 95–98
Appendix B: Biographical Sketches of Workshop Participants 99–122
Appendix C: Workshop Agenda 123–134
Appendix D: Registered Attendees 135–140
Appendix E: Technology Readiness Levels in the Department of Defense 141–144
Appendix F: White Paper 1: The BioWatch Program: What Information Is Needed to Inform Decision Making? 145–154
Appendix G: White Paper 2: Nucleic-Acid Signatures at Three Levels of Readiness for BioWatch 155–172
Appendix H: White Paper 3: State of the Art for Autonomous Detection Systems Using Immunoassays and Protein Signatures 173–196
Appendix I: White Paper 4: State of the Art for Autonomous Detection Systems Using Genomic Sequencing 197–214
Appendix J: White Paper 5: State of the Art for Autonomous Detection Systems Using Mass Spectrometry 215–244


Finance has at all times been a really busy and profitable sector which controls the economy of the world. Full entry to all the latest information coverage, with the flexibleness to view content material offline and save articles for later by a cell-optimized website and iOS app. It was acknowledged that the educating of financial accountability is and always has been a high priority for the Credit score Union League as a result of it's their knowledge that the financial future of the state is in jeopardy. If you can get your company to pay this for you as a business expense, it's a great publication to remain on high of financial information. Financial markets have began the month on an uncertain footing after a dramatic November, with the dollar retreating from a nine-month excessive. Trying to keep up with the Joneses, when the Joneses' real finances are unknown, is unwise, a financial planner reminds us.
However, it has been recognized that some WSJ articles are totally viewable for those who entry them through Google News. It's now online, but unlike many other media sites, the Monetary Occasions has chosen to cover behind a paywall. SmarterHQ will use these funds to broaden sales and advertising efforts as it continues growing into new vertical markets, including financial providers, travel and hospitality.
Hilton Worldwide (NYSE: HLT) in the present day announced the appointment of Thomas J. Baltimore, Jr. as president and chief govt officer and Sean M. Dell'Orto as chief financial officer of the planned real property funding belief (REIT) that's anticipated to type following its separation from Hilton Worldwide later this yr.
Lenders have raised interest rates on various monetary products, together with mortgages, loans, and bank cards, and have additionally tightened up on their lending criteria, leaving many shoppers out in the chilly in relation to getting finance.
Media coverage of the eruption brought elevated consciousness of the natural beauty of the distant nation which, when coupled with the severe decline in the Icelandic krona in comparison with other major currencies after the monetary crisis, led to an unforeseen explosion in tourism of volcanic proportions.


letter now through the help of my computer beside my sick bed.
This Email with the Subject "Urgent Respond from Mrs. lilian chen" was received in one of Scamdex's honeypot email accounts on Sat, 13 Aug 2011 03:20:07 -0700 and has been classified as a Generic Scam Email. The sender shows as lilianchenchenlilianch <lilianchenchenlilianchenchen@gmail.com>, although that address was probably spoofed. We recommend that you do not attempt to contact any persons or organizations referenced in this email, as you may expose yourself to scammers and, at the very least, you will be added to their email address lists for spam purposes.


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Home / News / Law & Government / Hochul prepares for spotlight as Cuomo steps down
Kathy Hochul/The Associated Press
Hochul prepares for spotlight as Cuomo steps down
By: The Associated Press, Adina Genn August 11, 2021	Comments Off on Hochul prepares for spotlight as Cuomo steps down
Kathy Hochul, a western New York Democrat, is known to Long Island business leaders as the state's lieutenant government. Still, many across the state are unfamiliar with Hochul, even after she's served in this role over the last six or so years.
On Wednesday, she began reintroducing herself to the public as she prepared to take the reins of power after Gov. Andrew Cuomo announced he would resign from office.
Hochul, 62, in less than two weeks will become the state's first female governor, following a remarkable transition period in which Cuomo has said he will stay on and work to ease her into a job that he dominated over his three terms in office.
She stayed out of public sight Tuesday but said in a statement that she was "prepared to lead." Hochul planned to hold her first news conference Wednesday afternoon at the State Capitol, in the very room where Cuomo became a familiar face to people across the U.S. and beyond for his televised briefings on New York's fight against the coronavirus pandemic.
Cuomo, 63, announced Tuesday that he would step down rather than face a likely impeachment trial over allegations that he sexually harassed at least 11 women, including one who accused him of groping her breast.
Cuomo has continued to deny that he touched anyone inappropriately, and said his instinct was to fight back against claims he felt were unfair or fabricated. But he said that with the state still in a pandemic crisis, it was best for him to step aside so the state's leaders could "get back to governing."
That job will fall to Hochul, who served briefly in Congress representing a Buffalo-area district, but purposely kept a modest profile as lieutenant governor in a state where Cuomo commanded — and demanded — the spotlight.
A seasoned veteran of retail politics, Hochul shares some of Cuomo's centrist politics, but is a stylistic contrast with a governor famous for his love of steamrolling opponents and holding grudges, state political veterans say. She's well-liked by colleagues, who say voters shouldn't confuse her quiet approach under Cuomo with a lack of confidence or competence.
"Lt. Gov. Kathy Hochul will be an extraordinary governor," Sen. Kirsten Gillibrand, another upstate political veteran, told reporters at the U.S. Capitol on Tuesday. "She understands the complexities and needs of our state, having been both a congresswoman and having been lieutenant governor for the last several years."
Long Island business leaders give Hochul high marks, noting that that she is well-versed on the challenges and strengths of the region.
Matt Cohen, president and CEO of Long Island Association, the region's largest business group, said that Hochul "has been a friend to Long Island during her entire tenure and has played a crucial role in supporting our region's economy, especially as we confronted the challenges with COVID-19, and we look forward to a continued relationship with her as our great State's first woman governor."
State Sen. Anna Kaplan, who chairs the Committee on Commerce, Economic Development and Small Business, said Hochul knows the region well.
Hochul "has often been called an honorary Long Islander because she spends so much time here fighting for issues that matter to us, and standing up for small businesses in our community," she said in a statement.
These are "extraordinary times," Kyle Strober, executive director of ABLI, said, and they
"require a steady hand and a clear vision of what the state needs – from public health to our economy."
Hochul, he said, "has demonstrated that strength of character during her tenure and the business community will welcome her presence in the governor's mansion."
Kevin Law said that he spent the last seven years working with Hochul in his role as co-chair of Long Island's Regional Economic Development Council.
"I have toured numerous businesses, vineyards and public works projects with her," he said . "She knows more about Long Island than most Long Islanders, and I am confident she will do a great job as governor and continue to be a supporter of our region."
The Retail, Wholesale and Department Store Union described Hochul as a "strong and smart leader with the right values," adding its expectations that she would "protect and advance the needs of working people during these extraordinary times."
It remains to be seen how involved Cuomo will be in state government over the next two weeks, or how he'll manage handing over authority — something he rarely ceded during his time in office.
His circle of advisers has shrunk, but his closest aide and policymaking partner — Melissa DeRosa — made a surprise return to Cuomo's side after having announced her resignation from the administration Sunday. The governor's office said she will remain in her job as secretary to the governor until Cuomo departs.
Leaders in the state legislature have yet to say whether they plan on dropping an impeachment investigation that has been ongoing since March, and which had been expected to conclude in the coming weeks.
In addition to examining his conduct with women, lawyers hired by the state Assembly had been investigating whether the administration' manipulated data on COVID-19 deaths in nursing homes and whether Cuomo improperly got help from his staff writing a book about the pandemic.
Republicans have urged the Democratic-controlled legislature to go ahead with impeachment, possibly to prevent Cuomo from running for office again.
As Assemblyman Jarett Gandolfo, a Republican from Bayport put it, put it, "Cuomo still has much to answer for."
Still, he said, that he "hope[s] to work together in a bipartisan fashion on behalf of all New Yorkers."
Andrew Cuomo Kathy Hochul 10:48 am Wed, August 11, 2021
Tagged with: Andrew Cuomo Kathy Hochul
Cuomo calls Trump tax plan a "scam"
Polimeni: P3s must offer better incentives to the private sector
LIers riled by state's clean energy tax
State eases insurance for domestic violence victims
Farmingdale State awarded $6.6M for transport security center

redpajama_set_name: RedPajamaCommonCrawl


text:
Q: I am trying to deploy my nextjs app to git hub page. but it is not deploying in the correct way I'm trying to deploy my nextjs app to git hub page. but the production is very slow. and everytime it publishes shows only the read.md. the way I deployed is below.

*

*installed the gh-pages into devDependencies.

*added replacer.js with following code..
const replace = require("replace-in-file");
    const options = {
      //you may need to modify the file address to suite your project
      files: "./build/index.html",
      from: [/src="\//g, /href="\//g],
      to: ['src="./', 'href="./'],
    };
    (async function () {
      try {
        const results = await replace(options);
        console.log("Replacement results:", results);
      } catch (error) {
        console.error("Error occurred:", error);
      }
    })();

*updated next.config.js is below
module.exports = {
      Images: {
        domains: ["/public", "fakestoreapi.com"],
        env: {
          PUBLIC_URL: "https://imangi.github.io/amazon-clone",
          assetPrefix: "./",
        },
      },
    };

*updated package.json file is as below.
{
  "name": "amazon-clone",
  "version": "1.0.0",
  "homepage": "https://imangi.github.io/amazon-clone",
  "description": "amazon clone with react & next.js",
  "main": "index.js",
  "scripts": {
    "replaceFilePaths": "node replacer.js",
    "test": "echo \"Error: no test specified\" && exit 1",
    "dev": "next",
    "build": "next build && next export -o build && npm run replaceFilePaths",
    "deploy": "gh-pages -d build",
    "predeploy": "npm run build",
    "export": "next export -o build && npm run replaceFilePaths",
    "start": "next start"
  },
  "author": "Imangi Nanayakkara",
  "license": "ISC",
  "dependencies": {
    "@heroicons/react": "^1.0.6",
    "@reduxjs/toolkit": "^1.8.2",
    "@tailwindcss/line-clamp": "^0.4.0",
    "firebase": "^9.8.2",
    "firebase-admin": "^10.2.0",
    "next": "^12.1.6",
    "next-sitemap": "^3.0.5",
    "react": "^18.1.0",
    "react-dom": "^18.1.0",
    "react-redux": "^8.0.2",
    "react-responsive-carousel": "^3.2.23"
  },
  "devDependencies": {
    "autoprefixer": "^10.4.7",
    "gh-pages": "^4.0.0",
    "postcss": "^8.4.14",
    "tailwindcss": "^3.1.2"
  }

}

*Executed the following command:
npm run deploy

*pushing _next folder with .nojekyll file to gh-pages branch.

redpajama_set_name: RedPajamaStackExchange


text:
Dunkirk screenings net £1,500 for Sidmouth RBL
Stephen Sumner
Published: 10:31 AM August 23, 2017 Updated: 6:57 PM November 7, 2020
Sidmouth Royal British Legion volunteers with staff from the Radway cinema - Credit: Archant
Military families will benefit from the £1,500 raised so far at the Sidmouth screenings of a blockbuster depicting the 'miraculous' evacuation of Allied soldiers in World War Two.
Radway cinema manager Amanda Marks invited volunteers from the town's Royal British Legion (RBL) branch to rattle their buckets after each showing of Dunkirk.
They were delighted to break the £1,000 mark and they will take the total higher still when the film returns for another week from today (Friday).
Amanda said: "It's all about supporting the community – we thank each and every customer for their donations to the RBL."
Radway deputy manager Caroline Vansittart said Dunkirk was busy for the three weeks it was showing and it is back due to popular demand. Screenings will begin at 8.20pm.
RBL branch chairman Dave O'Connor said: "We're very pleased with the sum they took – it's absolutely fantastic. Our thanks to the manageress and her staff."


text:
Q: GPS start mode query I am using SIM808 GPS/GSM module. I came across Cold start,Warm start and Hot start in GPS Modules while developing code for a GPRS Tracker for car. I know the difference between them.My question is that if I don't use any start mode then what will be the searching procedure of gps.

A: When the GPS receiver does a cold start, it's search space has three dimensions:

*

*test for each satellite to find which ones are actually in view

*Doppler shift

*pseudo random code offset

Any normal GPS module implements the search procedure internally.  You wouldn't need to worry about implementing your own search procedure.


Everyone is talking about it but what is it? I get asked alllllll the time, it seems to have captured people's curiosity. Well this paint has captured my heart! If you are not familiar with it and you've been wondering what it's all about carry on reading.
Chalk paint is not to be confused with chalkboard paint, which is used on a painted surface to create a chalkboard. Chalk paint can be used for a wide variety of painting projects, not just wooden furniture but metal and walls, just about anything you want to dream up. It leaves a wonderful matt finish and extremely forgiving to use.
Here at Sara Hughes Home I stock Autentico chalk based paints. They are water based, free of solvents and other harmful additives (VOC low) and can even be used in situations where anti allergic and bacterial features are paramount. People call it eco paints, organic paints, environmentally friendly paints, we call it Autentico!
Autentico chalk based paint was especially developed using old recipes and only natural ingredients. This information has been taken from their website, you can find further details about these gorgeous paints on my website under 'buy Autentico' or head to their website, www.autentico-chalk-paint.co.uk.
I'll tell you why I'm chalk paint mad. It's brilliant in the fact it can be applied in many different ways to create a variety of styles with various techniques such as textured surface, distressed finish, crackle it for an old world effect or just simply a beautiful chalky matt finish.
If like me you love giving furniture a new lease of life then chalk paint is your substance. Heaven in a pot. Give it a try!


text:
North West, UK & Irelandremove
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We found 6 jobs in North West, UK & Ireland
Click on the Details/Apply button next to each job to see the full posting.
Medical Sales Representative - Medical Physics
Area: North East, UK & Ireland - North West, UK & Ireland - Yorkshire & Humberside, UK & Ireland - East Midlands, UK & Ireland - West Midlands, UK & Ireland| Salary: £25,000 - £35,000 + £10,000 bonus, company car, benefits
As Medical Sales Representative for this global leader in medical dosimetry, radiotherapy and medical imaging QA & QC (quality testing) products, y...
Radiotherapy Service Engineer
Area: North East, UK & Ireland - North West, UK & Ireland - Yorkshire & Humberside, UK & Ireland - East Midlands, UK & Ireland - West Midlands, UK & Ireland - Wales – North, UK & Ireland| Salary: £45,000 - £55,000 + bonus, food allowance, car allowance, benefi
As a Field Service Engineer for this global company, you'll work with cutting-edge radiotherapy medical equipment, software and networking syst...
Sales Specialist - Ultrasound HLD
Area: North West, UK & Ireland - Yorkshire & Humberside, UK & Ireland - Wales – North, UK & Ireland| Salary: £50,000 - £60,000 + £25-30k OTE, car allowance, benefits
As a medical Sales Specialist for this global company, you'll sell ground-breaking Ultrasound Ultra-High Level Disinfection systems as part of a ...
Clinical Sales Specialist - Cardiology Devices
Area: North West, UK & Ireland - Wales – North, UK & Ireland| Salary: £35,000 - £45,000 + £20,000 bonus, car, benefits
As a Clinical Sales Specialist you'll promote and support Diagnostic & Invasive Cardiology equipment in hospitals across the North West as par...
Area: North East, UK & Ireland - North West, UK & Ireland - Yorkshire & Humberside, UK & Ireland - East Midlands, UK & Ireland - West Midlands, UK & Ireland| Salary: £45,000 - £55,000 + car/cash allow. + amazing benefits
As a Field Service Engineer for this global company, you'll work with cutting-edge Radiotherapy medical equipment in NHS and private hospitals....
Clinical Specialist - Spine Navigation & Robotics
Area: South East, UK & Ireland - South West, UK & Ireland - South Central, UK & Ireland - North West, UK & Ireland - Yorkshire & Humberside, UK & Ireland - East Midlands, UK & Ireland - West Midlands, UK & Ireland - London, UK & Ireland - Home Counties, UK & Ireland - East Anglia, UK & Ireland - UK Wide, UK & Ireland| Salary: £35,000 - £50,000 + bonus, car or cash, benefits
As a Clinical Specialist for this global image guided surgery and surgical robotics company, you'll support spine surgeons in operating theatres ...
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Born in 1981 in Delhi, Mohammed Zubair passed his graduation in arts from a Delhi University college, and holds a certificate in Arabic Language, and a diploma in Calligraphy and Graphic Design. His command over lines and letters coupled with his expertise in design and graphics is reflected in his calligraphic compositions in Arabic, Urdu, Hindi and English. Using a range of fonts and styles of calligraphy his art adorns, what he describes as Qalam-dawaat (wooden pen and Ink) marks. He has worked with Urdu Academy Delhi and with various NGOs. His calligraphic art has been part of several exhibitions and he has participated in and conducted various workshops including those organised by Indian National Ttrust for Art and Cultural Heritage, Dastkaar and National Mission for Manuscripts. Currently working as the Head Instructor at a calligraphy centre run by Delhi government, the artist is trying to keep alive a nearly extinct art form. Mohammed Zubair lives and works in Delhi.


Great Britain hotel takes its name from the English merchants and sailors who chose Siracuse as a meeting point for their trades, as well as an important passage to Malta.
Great Britain Hotel is an old hotel in Siracuse. It was been entirely restored in 2001 and is situated in the very hearth of the isle of Ortigia.
It is a few minutes walk from the monuments that make Ortigia one of the most beautiful places in the world for its unique mixture of ancient Greek ruins and Sicilian baroque.
The hotel overlooks the ancient ruins of Apollo Temple and is located a short distance from Piazza Duomo, where the splendid front of the baroque Cathedral, built on the ancient Athena Temple, can be admired and whose columns are still intact.
Not far from the Cathedral is the church of St. Lucy alla Badia and Senato Palace, a splendid example of late baroque, designed by Vermexio.
In Ortigia you'll spend a wonderful holiday bathing in its sea shores born along the coast or visiting the numerous pubs and bars that enlive the isle during the night. See you soon!


One of the more notable aspects of the current economic slowdown is the fact that many firms have become increasingly ''meaner'' to their associates. In fact, many firms have become so nasty that many associates often wonder why they chose careers as attorneys in the first place. We have heard stories that, quite frankly, are shocking the first time you hear them but now occur with such frequency they are becoming old news. For example, one major firm over the past year fired more than half of its first year associates class with little or no explanation at all. This firm simply informed these associates that it'' ''we do not believe you are a good fit. In itself such conclusions may not seem that terrible; however, when you consider the fact that these same firms never gave their associates a single performance review and offered no critique whatsoever of their work you quickly must realize that this is not the rational environment one expects from most organizations that employ highly educated professionals.
Not that all firms are mean, however, attorneys often need to be aware of the conditions within firms that can make them ''mean''. Indeed, an attorney's survival depends upon making choices of where to go to work based upon the perceived social and economic stability of the given firm. It goes without saying that these choices are some of the more significant choices attorneys need to make in order to survive.
As this article will demonstrate, firms that are doing well economically are often not mean, and it is important for attorneys to seek out firms that consistently perform well economically. Since law firms are by nature somewhat difficult to understand and not exactly forthright with information regarding their economic performance—some firms are often even described as portraying an ''aura'' of mystery—this is not always an easy thing to do. The best way to study this phenomenon is to contrast differing economic conditions at different points in time-in a good climate and in a poor one. For example, since 2000 there have been numerous associate layoffs throughout the United States. This contrasts with the massive hiring binge at firms in between 1998 and most of 2000. The way a firm treats is associates is generally a product of economic conditions affecting the firm.
Between 1998 and 2000, most attorneys who worked in firms which now have either closed or laid off a significant number of attorneys did not believe their firms were ''mean'' at all. In fact, in order to try and get associates to stay (so they could do all the work), firms tended to be extremely lenient in performance reviews. Interviews of potential hires were often quite casual and not too stressful. Bonuses were large. Firm perks were abundant. For many associates practicing at large firms, their biggest complaint was often that they had far too much work to do. Nevertheless, associates with strong credentials could easily switch firms if they thought their firm was not a pleasant place to work.
For associates working at "nice" firms between 1998 and 2000, they often believed they could do no wrong. When looking for positions, associates were often very arrogant about the number of opportunities they had. In associate interviews, many law students and lateral associates would often grill hiring partners about what the firm could do for them and not vice versa. Things were very good for associates.
In the Year 2001, many law firms became extremely unpleasant places to work. Performance reviews suddenly turned "nasty" when there had been no apparent change in the quality of the associates work. At many performance reviews associates were outright fired, or simply laid off. For corporate associates seeking to lateral into another law firm - in contrast to what occurred in the "boom years" - there were virtually no recruiter opportunities. With respect to the few interviews some of these associates received, the firms grilled them aggressively and were slow to hire. Bonuses? At many firms there were no bonuses. Perks were cancelled or dramatically scaled back. As for complaints about too much work, many associates were complaining about the exact opposite.
The obvious meaning of all this is that when economic times are good, associates make firms a great deal of money and are treated well. When economic times are bad, associates may cost their firms money and their careers will be in jeopardy. Accordingly, economic forces beyond an attorney's immediate control often shape their futures.
However, for all of the criticisms of firms during bad economic times, attorneys at all levels do have control over their destinies and how they can confront economic situations. For example, even during the worst economic times there are firms that treat associates well and post high profits. Many of these firms never turn mean. Even during recession periods, some firms are aggressively hiring attorneys. It is also the firms that grow during poor economic climates that are also most likely to offer the best future opportunities.
In law school, most students do not know what to look for in a firm. For example, during the boom years one of the most attractive law firms to many associates in the United States was Venture Law Group in Silicon Valley. This firm essentially took one aspect of a law firm's corporate department and built it into a law firm doing nothing but this certain type of corporate work. Conversely, a firm that has been around for over 100 years, such as New York's Sullivan & Cromwell, has survived so long because it has numerous departments that complement each other and help sustain it through economic cycles. Incidentally, Venture Law Group is a firm that "became mean" during the economic slowdown. According to what Legal Authority clients have said, this particular firm actually laid off several first year associates who were less than six months into the practice of law.
The essence of becoming a client of Legal Authority is that we give you the ability to distinguish the good firms from the mean ones which is crucially important in a bad economic climate. By allowing you to survey the market for all of the opportunities that match what you are seeking to do, Legal Authority empowers you to find firms that are actually growing in a bad economic climate. If you are a corporate associate, or practicing in an area of the law that is not in demand in a bad economic climate, finding firms that are hiring is an extremely significant thing: You have found a firm that is likely to be in even better shape when the economy turns around and where there will be opportunities.
How do you do this through Legal Authority? For one, most Legal Authority clients receive multiple offers. The number of offers you receive by using Legal Authority if a function over some forces we cannot control, such as your geographic location, practice area and particular skill set. Nevertheless, most Legal Authority clients do receive multiple offers because they are taking care of hitting every single potential job opening at one time. Through this process, Legal Authority clients are able to evaluate a number of offers at the same time. Once you have your offers in hand, the key is investigating the firms that are making the offers and asking questions of other associates in the firm. Most firms—if you ask—will welcome you asking people inside the firm about the firm once you receive the offer. Ask questions about turn over and the history of the firm in the present economic climate.
Another useful tactic is to examine many of the firm's clients. Most firms list their clients on their website and most have these clients listed in their marketing materials. A firm with a broad based client group and long-established clients is far more likely to have economic security. A firm with one major client is likely to be a risky place to work. A firm with clients all in one (economically vulnerable sector) is also likely to be a risky place to work. In addition, you should examine the firm's practice areas. If a firm has numerous practice areas and not a high proportion of attorneys in each they are likely to have more economic strength than if they rely on one practice area for most of their business. There are some exceptions to the later, however. For example, litigation is a practice area that can be strong in all economic climates. Nevertheless, if one firm has a high number of litigators due only to the fact that they are working on a major case such as a class action this might not necessarily be the best thing. Obviously, practice areas like corporate and bankruptcy are far more vulnerable to the economic climate.
Finally, you should trust your instincts about a given firm. How do the attorneys (and, most importantly, partners) in the firm make you feel. Do the attorneys seem content? Are they forthright and responsive to your questions? How do you feel when you are around these attorneys? What type of examples of success can they point to for individuals similarly situated to yourself who joined their firm. The list goes on and on. In the end, you need to realize that the economic health of the firm will have a serious impact on your economic security in the long run.
When firms turn mean, it is usually due to economic forces. Firms are set up in different ways and, as they evolve they make many economic decisions that have a significant effect on them in both good and bad economies. Finding firms that are set up to weather economic storms is no easy task. The firm could be a very large law firm, or it could as easily be a smaller one. Legal Authority clients are assisted in finding the opportunities that are set up to survive and even prosper in economic storms. By contacting all of the firms in a given city that match precisely what you are seeking to do, you can identify hiring firms and then speak with them and judge for yourself. While results differ for Legal Authority's clients, it is not uncommon for many of our clients to get more than 10 offers even in poor economies. This breadth of choice provides you with an unrivaled opportunities to make judgments and evaluations you might not otherwise have the chance to make.
So, You Want to Become a Law School Professor?


Are you ready for the 18th Annual Mardi Gras Texas Style! Festival at the Automobile Building in Fair Park? This Dallas staple will take place on February 23rd, 2019! Find outfits like this Over-the-top Mardi Gras Cowgirl Masquerade Costume at Dallas Vintage Shop!
This entry is filed under 2020 TBA (Q1) Mardi Gras Texas Style!. You can follow any responses to this entry through the RSS 2.0 feed. Both comments and pings are currently closed.


Within the team at Create Consulting there are a number of experienced expert witnesses.
Jonathan Cage, the Managing Director, specialises in expert witness in highways, transport and general infrastructure provision issues. Jonathan has previously given evidence at numerous planning appeals, CPO Inquiries, examinations in Public, Mediation Sessions and is used to working with Queen's Counsel and leading Barristers.
Mark Allen acts as an expert witness within the Highway and Transportation, Mark has prepared and given evidence at planning inquiries.


These files are for Educational Purposes ONLY! I make no claim about their performance/operation/origin. Please don't blame me if you use something here and it doesn't work or breaks something.
This project started because I found out a buggy EPROM in my '85 TPI Z28 was causing a hesitation in 3rd-gear@3600rpm. I'm trying to collect/archive stock GM/FBody EPROM images so that people have access to the stock code if they suspect their EPROMs are causing them problems, or if you're swapping engines/ECMs in your GM/FBody.
If you have the means to copy your Stock EPROM , please email it to me to include it on the list below. If you have your stock EPROM out of the car (replaced with an aftermarket?) and can Snail-Mail it to me, I can read it and send it right back.
GOTO -> Nat. Hwy. Trans. Safety Admin. Lots of info on TSBs/Recalls/etc.
(Origin Unknown, most parameters same as 85vette2) ?
85-mpfi-V6.bin '85 Camaro MPFI ????
86TPI.bin '86 TPI (305?) ????
87GTA350.bin '87 GTA 350 ????
88camaroTBI.bin '88 Camaro TBI ????
90camaro350.bin '90 camaro 350 ????
90-350(L98)-M5.bin '90 350 (L98) w/M5 ????
91GTA305A4.bin '91 GTA 305 w/A4 ????
92A4.bin '92 Fbody w/A4 ????


The Technology Investigation team is working on exciting new technologies and we are looking for a dedicated Embedded Systems Software engineer. As a member of our fast-paced group, you will have a unique and rewarding opportunity to be part of a new project and shape upcoming products that will delight and inspire millions of Apple's customers every day.
Experience developing real time software for multi-sensor systems.
Expertise in low level image, vision and signal processing pipelines.
Expertise developing drivers and firmware to accelerate performance of real time media applications. Knowledge of low level hardware and OS internals at a kernel level.
Experience with Rapid Prototyping, POC development and debugging.
Curious and loves working with the best creative people in the industry to solve hard technical problems.
Passionate, Inspired by Apple's mission and products.
You will work closely with software, hardware, algorithms and UX teams to create ground breaking technologies and systems. You will architect and develop high quality software for real time interactive media applications. Your knowledge and experience in the area of low level software development and hardware acceleration for video, image processing, computer vision will be leveraged. You are curious and love working with the best creative people in the industry to solve hard technical problems. Grit and ability to persevere are key.


text: Q: Why is my heading and text not appearing? I'm sure this is really simple, sorry but I'm a web noob. I can't get the headings to show up on the page.


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Jonathan Kraft On Ravens Struggles: 'It's Too Bad, Isn't It?'
Filed Under:98.5 The Sports Hub, Baltimore Ravens, Jonathan Kraft, New England Patriots, NFL, Sports News
Baltimore Ravens head coach John Harbaugh. (Photo by Ezra Shaw/Getty Images)
BOSTON (CBS) –The Baltimore Ravens are off to an 0-2 start to the season, but they won't be getting any sympathy cards from the New England Patriots.
"It's too bad about Baltimore, isn't it?" Patriots president Jonathan Kraft said to Marc Bertrand on 98.5 The Sports Hub Sunday morning, ahead of New England's game against the Jacksonville Jaguars. "It's really too bad about Baltimore."
When asked if Baltimore's struggles may be a little bit of DeflateGate karma coming back to haunt them, Kraft wouldn't bite.
"That has nothing to do with it," he said. "The Ravens are sweethearts. [Head coach] John Harbaugh, he's a sweetheart."
It's a shame these two teams aren't scheduled to face each other in the regular season.
Listen to the full pregame interview here:


Whenever I prepare this mung dhal the smell of sizzling spices teleports me to Hassan's kitchen in Madame Mallory and the smell of curry. It's irresistible.
Soak the mung beans during the night or for a minimum of 4 hours.
Bring the water to boil in a saucepan, add the mung beans, the curry powder and the turmeric. Simmer on medium heat with a lid on. Cook for 25 minutes. Finally stir in the rock salt.


Destination weddings are some of the most memorable events for everyone involved. Whisking your close friends and family away to spend time together and celebrate your marriage creates a shared experience that can't be replicated. At Florblanca, we have an exotic location and intimate setting that's perfect for destination weddings.
We recently hosted Megan and Trent's wedding and it was an absolute dream. They had such a gorgeous vision for their celebration and we're thrilled that we were able to meet their expectations.
Bliss Weddings, a wedding planner in Santa Teresa, shared photos of this event from Sara Ford Photography.
We just had the most magical week of our life celebrating our wedding at Florblanca. We literally could not have imagined a better experience for the whole week, and especially for the wedding. The location, the resort, and especially the staff went above and beyond to make it literally the best wedding of all time!
Thank you to Megan and Trent for trusting us with this incredibly special event. We are so happy for you and wish you the very best always.
Learn more about Weddings at Florblanca.


text:
From Whistler to Warhol: Modernism on Paper
Home / Works of Art / Exhibitions / Previous / From Whistler to Warhol…
Jackson Pollock, United States, 1912-1956, Untitled, 1951, Screenprint, Edition of 25, Gift-Purchase. Craig F. Starr Associates and the John Levas Fund, 2009 (31434),© 2009 The Pollock-Krasner Foundation/Artists Rights Society (ARS), New York.
Joseph Stella, Born Italy, Active United States, 1877-1946, Abstraction, 1919, Pastel on paper, Purchase. The John Levas Fund, 2009 (31433)
February 18, 2010 - July 03, 2010
Henry R. Luce Gallery (28)
Exhibition Overview
When Anna Rice Cooke founded the Honolulu Academy of Arts in 1927, she donated a core group of works of extraordinary quality whose variety reflected her mission to present art resonant with the diversity of modern Hawaii. Included in this inaugural gift were numerous European and American prints and drawings, for work on paper was an integral part of the museum's original program. Over the years, the Academy's prints and drawings collection has grown to over 15,000 objects, most of which date from the mid-19th and 20th centuries. In an unprecedented move, the Academy presents a major exhibition of 100 masterworks from these modernist holdings, including a number of unpublished and rarely exhibited examples, as well as a selection of important recent acquisitions.
Beginning with the American painter-printmaker James McNeill Whistler, the exhibition traces the history of modernism on paper to show how this technically variable, readily portable, and easily disseminated medium proved supremely adaptable to the avant-garde spirit of experimentation. It shows how Whistler pushed the limits of etching to elevate its status to that of painting, and how his lead was followed by many of his European contemporaries—among them the impressionists Camille Pissarro and Edgar Degas; and the symbolists Odilon Redon, Edvard Munch, and Paul Gauguin—all of whom likewise took printmaking beyond its illustrative function. They also asserted drawing as a primary medium, establishing the graphic arts—printing and drawing—as independent modes of expression.
The exhibition continues into the early 20th century, when the cubists Pablo Picasso and Georges Braque, the German expressionists Emile Nolde and Käthe Kollwitz, the surrealists Max Ernst and Joan Miró, and the American modernists John Marin and Joseph Stella exploited the blank page as a tabula rasa for new ideas both artistic and political. Special focus is given to American printmaking of the first half of the 20th century, when artists such as Edward Hopper, John Sloan, Raphael Soyer, and Thomas Hart Benton mastered etching and lithography as vehicles for conveying their fascination with the dynamism of the city and the quiet toil of rural life.
The exhibition continues into the mid-20th century, when the realism of the American Scene was supplanted by the abstraction of the internationalist New York School, whose revolution in painting found a counterpart in the innovations in printmaking achieved by many Abstract Expressionist artists. And it culminates with the 1960s and 1970s, when master printers such as Tatanya Grossman and June Wayne set up collaborative workshops where artists such as Jasper Johns, Ellsworth Kelly, Roy Lichtenstein, Robert Rauschenberg, and Frank Stella created screenprints and lithographs that took printmaking into the largely uncharted territory of photomechanic processes. Many of these artists—among them Sam Francis and Andy Warhol—set up print workshops of their own, asserting the centrality of printmaking to their studio practice.
The exhibition is organized by the Honolulu Academy of Arts and curated by Theresa Papanikolas, curator of European and American art.
In the House: Recent Acquisitions
Constellation—drawing in space by Marian Bijlenga
Hayv Kahraman: Superfluous Bodies
Melissa Chimera: Migrant


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Q: Install Linux Mint External Hard Drive Partition I am currently trying to install the new Linux Mint 18 on an (empty) Seagate Expansion 1TB external hard drive. I am running Linux live from a USB and using the "Install Linux Mint" wizard that is included with the distribution.
I created 4 partitions:

*

*50GB "/" partition

*500GB "/Home" partition

*50GB "SWAP" partition;

*20GB EFI System partition.

However, when I try to install, I get the message:

The partition /dev/sda1 assigned to / starts at an offset of 3584 bytes from the minimum alignment for this disk, which may lead to very poor performance."

For each partition, I am placing everything at the beginning rather than the end. When I try to delete the new partition and recreate it, there is the same around 33MB of free space at the beginning and the wizard will not let me continue to the next step until I get rid of it.
Does anyone know what I am doing wrong and if there is a way to put the root directory at the beginning of the external hard drive and get rid of the free space?

A: I was having the same problem. I did about 5hrs of research on the net and many people had the same problem with the seagate external drives. It must be the usb interface causing the error. i took the drive out of the case and installed it in my e machines desktop and installed the os that way. When done i reinstalled the drive back into the enclosure, hooked it up to my HP G6 laptop which has no internal hard drive, set the bios to boot from the seagate backup plus and it worked. That was the only way i could get it to install.The computer that is using the external HD is currently doing updates.

A: FYI, it can be a problem installing Linux Mint 17.3 on "newer" (post-2009) SATA HDD that are 250 GB or above in capacity because of newer HDD technology = partitions not properly aligned with the disk. Something to do with 512 bytes data blocks in older IDE/PATA HDD (that came in 120 GB or less sizes) and 4096 bytes data blocks in newer HDD. E.g., the error message during installation = "... offset of 3584 bytes from minimum alignment ...".
If you get this error message during install of Linux Mint 17.3 on an external/USB HDD, you need to first use GParted (which is included in the Live DVD or USB-stick) to realign the modern SATA hard disk, i.e., unmount the HDD (usually is /dev/sdb or /dev/sdc), delete the whole old partition and create the first new partition meant for the "/" or root partition (e.g., 20 GB in size), with 1 MiB (= the default value) at the "free space preceding" and 0 MiB at the "free space following", ensure "MiB" alignment box is checked (= do not select "cylinder"), set it as Primary partition and format to FAT32 file system (= quicker process, i.e., no need to use ext4 yet).
....... When creating the new 2nd partition meant for the home partition (e.g., 30 GB in size), set 0 MiB for both the free space preceding n following, Primary partition and format to fat32.
....... If you have less than 4 GB of RAM, you can also create a new third partition for swap area (e.g., set as 2 GB in size) for virtual memory/RAM on the hard disk, ... do as for the home partition above.
You can leave the remaining space as unallocated or free space. After the install and boot, you can create a fourth new partition to store your back-ups, movies, music, photos, files or install another Linux distro (= need to sudo update-grub to dual-boot).
Only after partitioning the external HDD with GParted, can you proceed with the Install via the Live DVD or USB-stick. Click "Something else", click the first partition that you had created earlier with GParted, click "Change", click "ext4 file system", click "/" for Mount point, and so on, and also for the /home and swap partitions.
....... Ensure that the device for the bootloader installation is set to the external HDD, e.g., /dev/sdb or /dev/sdc. The cptr's internal HDD/SSD is usually identified as /dev/sda.
Seems the Linux installer does not realign modern SATA HDD when you create a "New Partition Table" for the / (root), /home and swap partitions during the install process.
I have just successfully installed and booted LM 17.3 on a "modern" SATA 250GB external/USB HDD which initially had the above error message about the first root partition not properly aligned with the HDD during install = could not proceed with the install. This was after many hours of reinstalling, trial and error.
....... Previously, I had no problem installing and booting LM 17.3 on an old external IDE/PATA 80GB HDD, i.e., no need to use GParted.


Show Me the #STEM Money!
Earlier this month, the Trump Administration announced its intent to bolster STEM education in the United States by offering new dollars for computer science education. But at first blush, it looks like an effort to throw pennies at an issue that deserves dollars (and a real commitment).
Over at TrumpED on the BAM! Radio Network, we explore the topic and what it really means for the future of STEM. Give it a listen!


The energies of spring are always alive within us regardless of the time of year. These energies represent all things new, naïve, innocent, and fresh. If you're into Tarot, it can be thought of as the Fool in the Major Arcana. I particularly love these energies when it comes to accessing and repairing the relationship with my inner maiden, or inner child.
As I've discussed before, we ALL have both masculine and feminine attributes; we are all everything and no-thing at the same time. If a gender specific sounding concept like "maiden" isn't resonating with you, think of this as our inner child, our inner innocence and purity, our inner clean slate – before life got a hold of us.
Aside from slightly longer and warmer days, what does spring actually bring and what are the sacred medicines and invitations of this time – AND what does it have to do with our inner littles?
Spring holds the energies of birth, rebirth, resurrection, beginnings, air, insight, the mind, divine inspiration, dawn, hope, promise, youth, beginnings. This is a time of year for emerging from our cocoons and slumbers and being with the light, the warmth, the potency and fertility of life, the place of endless potentiality and possibilities. It's a time to deeply connect with spontaneity, to release limiting viewpoints and beliefs and work with the mind and the element of air.
After the heaviness of winter, in all senses, we must bring movement back into our lives. What does this need to like for you right now?
Who is the Inner Maiden?
Our inner child holds our soul's mission and knows our highest passions in life. She knows our deepest desires and needs, our innocence, our joys, our reason for incarnating as a human.
We all came here with a specific mission – specific lessons to learn, things to experience, and some unique way to contribute to the healing of the collective. Spring is a time of resurrection and so it is the best time to resurrect the maiden and to come into right relationship with her and all of her splendor. She doesn't need to be a severed piece or a traumatized piece that is running our life. We don't need to view the world through trauma-tinted glasses, and we don't need to operate without the guidance of this all-too-important piece of ourselves.
Ever notice how we have this need to prove ourselves right and have the world fit into our narrow definition of it? This is the time to broaden that perspective and allow the potent element of air and the winds of spring to expand and bring with them spontaneous insights. I love wind – it's incredibly magical to me. I have such an affinity for the air element. It's ancient and has always been here and carries with it the whispers and secrets of those long, long ago. It carries divine inspiration and insight and can expand and permeate into every facet and nook and cranny of your being. It feels especially primordial to me and innocent and like, the basis for all else.
So, the inner child is meant to greatly influence our life because she knows why we are here, however, like we've established, she is also the one who remembers and experienced firsthand our earliest traumas and woundings and conditionings and as such, she decided a long time ago that it wasn't safe to be in the world as a sensitive, intuitive, empathic, spiritual being. She pledged to survive and stay safe and so she adopted many coping mechanisms and armors and filters and beliefs and personas and operates from them through adult you today… essentially a traumatized child is running your life. And in this way, we can forget why we're here, what we love, and what brings us joy because we're constantly seeking safety through perpetuating our narrow, traumatized view of the world.
There are ways to repair the connection and soften the trauma so we can begin to find stability in ourselves to extract the wisdom and gifts from these early events and then feel our subtle energies, which guide us towards our pleasures and joys.
• Coming into a state of body trust through practices like embodiment, yoga, supportive nourishment, herbalism, and more support the resurrection of the maiden.
• Aligning with Nature and seasonal flow and seasonal living support the resurrection of the maiden.
• Leveraging our subtle energies through shadow work, energy healing, flower essences, and more are probably the most important factors in resurrecting the maiden.
• Additionally, consciously hold space and ceremony to invite her into your life. Ask her what she wants to do, think back to what you enjoyed as a child, invite more play and lightness into your life. Essentially all things spring!
When do you remember "growing up?" When do you remember tender innocence? How can you merge these parts of yourself for a more cohesive, integrated experience? What would life be like if you were able to hold both your maiden self and your adult self? Our inner child can't do it all alone and clearly our adult selves without the input of a healthy inner child are only half as powerful… Allow these two to come together and create a third energy. Allow the potency of spring and the expansive nature of the air element support you always – regardless of the time of year! Here's to your beautiful maiden and your continued emergence, sensitive and magical soul!