text:
A linear low dropout voltage regulator is described that makes use of a depletion mode NMOS pass transistor and of a PMOS transistor in series to the NMOS transistor and connected to its drain. The depletion NMOS transistor assures low dropout operations, while the series PMOS transistor allows the current regulation even under the condition of shorted load. The same PMOS transistor may be used to disable the current in the load without generating a negative voltage at the gate of the depletion pass transistor. This regulator is inherently stable without the need for an output capacitor in parallel to the load.
1. A linear voltage regulator comprising:
a transistor having a drain, a source, and a gate, said source being electrically coupled to a load at a regulated voltage, said gate being coupled to a signal for controlling the voltage at said regulated load; and
a circuit for regulating the current in said transistor coupled to said drain of said transistor.
2. The circuit of
3. The circuit of
4. The circuit of
[0001] The present application claims priority from U.S. Provisional Patent Application No. 60/409,040 for LOW DROPOUT VOLTAGE REGULATOR USING A DEPLETION PASS TRANSISTOR filed on Sep. 9 2002.
[0002] 1. Field of the Invention
[0003] The present invention is in the field of electronic circuits. The present invention is further in the field of analog integrated circuits. The implementation is not limited to a specific technology (i.e. CMOS or bipolar), and applies to either the invention as an individual component or to inclusion of the present invention within larger systems which may be combined into a larger integrated circuit.
[0004] The invention also falls within the field of DC voltage regulators and electronic power supplies, which convert energy from one DC level to another. These devices have been common in all electronic systems. More specifically, the invention falls into the class of voltage regulators referred to as series pass regulators or low dropout regulators, which convert a higher voltage to a lower voltage.
[0005] 2. Brief Description of Related Art
[0006] Integrated circuit voltage regulators are common components which typically have an input terminal for receiving an input voltage, a common (ground) terminal, and an output terminal which supplies current to a load. The output terminal provides a substantially fixed voltage independent of the magnitude of the input voltage or the current provided to a load, provided that the input voltage is greater in magnitude than the desired output voltage.
[0007] Although many integrated circuit regulators provide this function only, it is common to provide additional functions in order to protect the circuitry and/or the load. It is usual to provide a mechanism to limit the maximum current the regulator will present the load. Many regulators also provide a means for disabling the output current, allowing an external enable/disable signal to determine whether the load will be powered. This is typical in large electronic systems with many individual functional blocks, where it may be desirable to selectively turn off those blocks to reduce power consumption when they are not required. Additional protections, such as over-temperature shutdown, are also common.
[0008] Available regulators can be characterized as either shunt regulators, which place a dissipative element in parallel with the load and control the shunted current to control the output voltage, or series pass regulators, which place a dissipative control element directly between the input voltage and the load. The latter technique has the advantage of being significantly more efficient than the shunt variety, and is the dominant approach used among integrated circuit regulators, and is the technique used in the present invention.
[0009] Among series pass regulators, there are two general classes. Conventional regulators use series pass elements which are unity gain followers (emitter followers or source followers), typically NPN or NMOS devices. This class of conventional regulator, in its integrated circuit form, is well described in Widlar. In order to drive the base or gate terminals, respectively, of these devices, the controlling signal must be higher in magnitude than the output voltage. This control signal requirement limits the �dropout voltage�, the difference between the input and output voltage of the regulator. In order to remove this limitation, a class of devices referred to as �LDO� or Low DropOut regulators was developed which used common emitter or common source output stages, typically PNP or PMOS transistors. The prior art circuit 1 using the PMOS transistor is shown in FIG. 1. Because the control signal (base or gate voltage) of these devices swings negative with respect to the emitter or source terminals, this control signal is not limited by the input voltage and it is possible to operate these devices with extremely small differences between input and output voltages.
[0010] Although the low dropout of the standard LDO circuits is very desirable, this architecture has some severe limitations in performance. The conventional regulator (using NPN or NMOS pass transistor) typically has much lower output impedance. The LDO typically requires a large capacitor at the output to maintain stable operations. Many LDOs are sensitive not only to the magnitude of capacitance across the load, but also to whether that capacitor looks like an ideal capacitor or whether it has a series resistive component at high frequencies. Selecting the wrong capacitor (too large or too small, too much series resistance or too little) can cause the LDO to oscillate.
[0011] The overall architecture of the series pass regulator is typically that of a feedback amplifier (as disclosed in the book �Analog Devices� in the chapter �Low-Dropout Regulators� by W. Jung). As shown in FIG. 1 such a regulator 1 includes an error amplifier A1 having an output connected to a gate terminal of the power transistor M1. A reference voltage generator V2 is amplified by a high gain feedback amplifier. As with all feedback systems, the performance is improved by increasing gain, but with a requirement that gain be rolled off at high frequencies in order to maintain the stability of the feedback loop. The mechanism for so limiting the high frequency gain is referred to as �compensation� and is of key importance in the design of all feedback systems.
[0012] In conventional regulator systems using unity gain follower outputs, the typical stabilization mechanism is to use a three stage amplifier. The first stage is a fixed transconductance, the second is a voltage gain stage, typically very high gain, which then drives a unity gain follower output stage. A feedback capacitor from the output or from the input to the follower, or both, is connected back to the output of the transconductance stage. This feedback around causes a dominant low frequency pole. This architecture is identical to the traditional feedback used in operational amplifiers.
[0013] Because this architecture has inherently low output impedance, which is further lowered by feedback, the system is relatively insensitive to loading. The reduction in feedback with increasing frequency can make the effective output impedance rise with frequency, causing it to look inductive. This inductive output impedance can, under certain circumstances, interact with capacitive loading to reduce the stability of the system, but the systems are generally very wideband and load insensitive.
[0014] The standard LDO is quite different in its frequency compensation. Typically the amplifier has two or three stages. An input stage compares a measure of the output voltage to the voltage reference. This stage may drive intervening stages, but eventually controls the common source/emitter output device. That final power stage provides voltage gain as a function of its transconductance and the load impedance (Av=gm*ZL). Since the load typically includes a capacitive component, that capacitor can be used to provide some of the gain reduction at high frequencies needed for stability. But typically the load capacitance is controlled by system requirements other than optimizing the stability of the LDO. It is therefore desirable to make the LDO stable over a wide range of capacitances.
[0015] It is not possible to use existing commercial LDOs without a large capacitive load (equal to or exceeding 1 uF). This results in the control loops of most LDOs being relatively slow. Since the LDO has very high output impedance without feedback, and a relatively low gain at high frequencies, it cannot maintain its output voltage in the presence of fast load changes.
[0016] To date, the primary approach to reduce the output capacitance sensitivity of the LDO has been to optimize the frequency compensation. Miranda (U.S. Pat. No. 5,686,821) and Brokaw (U.S. Pat. No. 5,631,598) use local capacitive feedback around the output devices and the driver stages to make these stages behave in a manner more similar to conventional output circuits using followers. Bakker et.al (U.S. Pat. No. 6,373,233) provided a somewhat similar solution, using a distributed RC network or its lumped equivalent around the output device alone.
[0017] Castelli et. al (U.S. Pat. No. 6,300,749) introduced a solution to add a mobile zero in the compensation circuit that is dependent on the second output pole of the LDO.
[0018] In all these cases the disadvantage is the need for an output capacitor to guarantee stability and adequate filtering of the output voltage.
[0019] There have been limited attempts to directly implement the older, faster control scheme in LDOs. One means of doing so is implemented in the UC385 regulator from Unitrode (now Texas Instruments). This regulator, element 2 in FIG. 2A, requires the introduction of a second higher voltage supply voltage V3 from which to run the control circuit. Power flows from the input supply V1 to the load with very low dropout voltage, but the gate/base drive of the pass transistor M2 is generated from the higher voltage supply. In principle, this second, higher voltage supply could be generated by the regulator 3 using a means such as a charge pump 4, as shown in FIG. 2B, but this would create unwanted noise and would delay startup until this required rail is generated. Such a regulator was introduced by Burr-Brown (now Texas Instruments), the REG101 and more recently by Philips, the SA57000-XX.
[0020] A more useful approach is the application of depletion mode power devices as pass transistors. Depletion mode devices are those where the turn-on threshold of the device is of a magnitude that zero control voltage allows the device to be conducting. JFETs and vacuum tube devices are inherently depletion mode devices, whereas bipolar transistors are inherently enhancement mode devices, inherently �off� with their control (base) pin held at the same potential as the emitter. MOSFETs can be made either enhancement or depletion by adjusting the surface concentration of the channel region. Most production CMOS processes include ion implantation steps to adjust the threshold of NMOS and PMOS devices to a desired threshold, typically a fraction of a volt. But an additional selective implant into devices destined to be depletion FETs can easily alter the threshold such that it is negative, forming depletion devices. This allows a standard CMOS process, with one additional mask step, to include depletion mode devices. Any process flow that builds enhancement mode MOSFETs can be modified slightly to provide depletion mode devices.
[0021] Wrathall et. al (U.S. Pat. No. 5,506,496) is an example of the use of depletion mode MOSFETs. There are several problems with the use of depletion mode pass devices, which are normally �on� and must have a negative voltage applied to their control terminal to turn them off. One problem is that under a condition of shorted load, where the output is at ground potential, the device will be on and cannot be turned off without the application of a negative gate voltage. Another potential problem with using depletion mode devices is that they are uncontrolled when voltage is initially applied. This causes the output voltage to be identical to the input voltage at start-up. Only after sufficient voltage exists to hold the gate below the source (output) by a voltage greater than the threshold voltage of the FET can any measure of control be imposed.
[0022] Wrathall's solution, to both problems, shown in FIG. 3, element 4, was a regulator 4 with PMOS device M3 as a switch in series with the source of the depletion mode MOSFET MD1.A switch control circuit 5 can selectively turn off PMOS device M3 in order to turn off current to the load. This implementation is not the ideal configuration, because in the condition of very low dropout voltage, it is necessary to fully enhance both NMOS and PMOS devices, i.e. maximizing the voltage from gate to source. When the regulated voltage is low, the PMOS device M3 in Wrathall cannot be fully enhanced. Similarly, by tying the source of the NMOS MD1 to the source of the PMOS M3, the Vgs which can be applied to the depletion NMOS is reduced resulting in increased total on resistance or bigger die area.
[0023] An earlier precedent for using �normally-on� devices comes from early regulator designs using thermionic devices (vacuum tube triodes and beam power pentodes). Vacuum tubes, like modern depletion FETs, were normally on with their control terminal (grid) held at the cathode voltage. By pulling the grid negative, the device could be turned off. A 1954 circuit for the HP 712B power supply, depicted in FIG. 4, shows very similar architecture to the conventional solid state series pass regulators discussed here. The gas discharge device U8 provides a voltage reference, a feedback amplifier comprising four triodes U1 through U4 compares this reference to a voltage divider taken from the output, and the amplifier drives a pass device U5, a beam power pentode. Note that as in the LDOs formed from enhancement devices in FIG. 2, this circuit requires a multiplicity of bias voltages, V5 and V6, in addition to the primary input voltage V4. Like the regulator using depletion MOSFET devices as in FIG. 3, this circuit required a switch SW1 that could keep the load disconnected during start-up, as the output voltage could rise to an uncontrolled high voltage before the active circuit could control it.
[0024] Accordingly, what is needed is a low dropout voltage regulator that combines the features of inherent stability, the ability to turn on and off very swiftly, the possibility to include a reliable means for limiting the output current and more importantly the capability to react extremely quickly to a change in load conditions. This would allow operation without the need for the output capacitor to filter the output voltage spikes and to provide stability to the control loop.
[0025] The present invention provides a fast LDO regulator which is insensitive to capacitive loads. This insensitivity allows the LDO to be used without requiring a capacitive load or, if a capacitive load is used, without imposing requirements on the value or quality of that capacitor. The fact that the LDO may be used without requiring an output capacitor, in some applications where it is required to turn off and on the regulator often to save energy stored in the batteries, such as in cellular phones, is a significant advantage because the energy stored in the output capacitor during the on time, is then left in the capacitor at the turn off. If the off time is long enough, due to the natural current leakage present in any capacitor, the capacitor discharges itself, resulting in energy wasted at every cycle. In addition the removal of the output capacitor improves the reliability of the overall system and reduces substantially the physical size and the system cost.
[0026] Because of its high speed, this present invention improves significantly upon the precision of the output in the presence of fast transients changes in the load current. One of the advantages of the described configuration is the fact that the higher intrinsic stability and better frequency response allows a potentially higher DC gain resulting in a much better load regulation with respect to a more traditional low drop-out linear regulator.
[0027] Furthermore in a configuration where the back gate of the depletion transistor is tied to the substrate of the IC (most common configuration of CMOS processes) the intrinsic body diode between input and output is eliminated and this could be advantageous in some applications.
[0028] A simple implantation allows the addition of a depletion transistor to any CMOS process without increasing the overall cost of the regulator.
[0029] The most general embodiment for the low dropout voltage regulator using the depletion type field effect transistor as main pass element is shown in FIG. 5.
[0030] The linear regulator 6 comprises a voltage control circuit 7 to control the voltage at the gate of the transistor MD1 in order to regulate the voltage at the load.
[0031] Furthermore a current control circuit 8 controls the voltage applied to the gate of PMOS device MP1 in order to control the current to the load.
[0032] According to the embodiment of the present invention, the depletion pass transistor MD1 is configured as a follower to allow the gate voltage to regulate the voltage at its source. Its back gate could be shorted to the source, but in a more common embodiment is connected to the substrate of the device.
[0033] The PMOS MP1 connected in series to the drain of MD1 allows for a complete shutdown of the regulator that otherwise would not be possible due to the negative threshold voltage of MD1. Furthermore MP1 could be regulated linearly to control accurately the current in the load providing a current limit function. This current limit could be a fixed one or could also be made a function of the output voltage as used in techniques referred to as �fold-back� current control.
[0034] According to the general embodiment of the present invention as shown in FIG. 6, a low dropout regulator with an input terminal Vin and output terminal Vout is provided consisting of a voltage reference V2, a differential error amplifier A1 comparing said reference to a measure of the output voltage, and a depletion mode FET MD1 which has its gate driven by the error amplifier output and its source tied to the Vout terminal. The drain of the depletion mode FET MD1 is connected to the drain of a PMOS MP1. The PMOS transistor has its source connected to the input terminal Vin and its gate tied to a control generator of function to be described, which under normal operation holds the PMOS switch in an �on� state.
[0035] In typical operation, the present invention operates similarly to the regulators described above of conventional design, but with the low dropout capability of an LDO. The use of a depletion mode device as a pass element removes the requirement of an input voltage which is substantially greater than the desired output voltage.
[0036] In normal operation, the PMOS MP1 switch is fully enhanced. This placement of the PMOS device has significant advantage over prior art Wrathall. When the regulator is being operated with substantial voltage between input and output, this configuration provides the benefit that the PMOS resistance is in the drain circuit of the NMOS pass device, rather than in series with the source. This allows for lower open loop output impedance, which improves performance.
[0037] The control circuit for the PMOS MP1 additionally is used to control fault conditions. In the case of a shorted load, where the output terminal is at ground potential, it is not possible to drive the gate of the depletion device below ground to reduce the output current. Under this condition, the PMOS can be programmed to operate at a fixed current which will control the current through the depletion NMOS or with a current dependent on the regulated output voltage providing the benefits of current fold-back technique to limit the power in the pass transistor in case of shorted load. In addition, it is possible to turn the PMOS MP1 transistor off to provide a �shutdown� mode where the LDO provides no current in the load. This shutdown mode may either be contingent on a fault (such as temperature exceeding a fixed threshold or input voltage exceeding a threshold) or it may be used to provide a system-level control of power to the load.
[0038] In a preferred embodiment of the present invention as shown in FIG. 7, the operational amplifier A2 drives the gate of the PMOS transistor MP3, which in its turn controls the voltage at the gate of the PMOS transistors MP1 and MP2.
[0039] In further embodiment of the present invention as shown in FIG. 8, the error amplifier and the reference voltage generator are combined in the classical Bandgap circuit 10 comprising transistors Q1, Q2, Q3 and Q4.
[0040] A further embodiment of the present invention shown in FIG. 9 presents a different type of voltage reference for use with CMOS process technologies. In addition an operational amplifier A3 acts as a voltage shifter to generate a replica of the voltage at the drain of MP3 to the gate of the transistors MP1 and MP2.
[0041] Further details of the present invention are explained with the help of the attached drawings in which:
[0042]FIG. 1 is a circuit diagram showing the prior art of PMOS Low DropOut voltage regulator;
[0043]FIG. 2A is a circuit diagram showing a prior art NMOS LDO regulator implemented with the use of an external higher voltage source generator to drive the gate of the pass transistor;
[0044]FIG. 2B is a circuit diagram showing a prior art NMOS LDO regulator implemented with a charge pump circuit to generate a voltage drive for the gate of the pass transistor;
[0045]FIG. 3 is a circuit diagram showing a prior art depletion NMOS LDO regulator implemented with a PMOS switch in series to the source of the NMOS as in Wrathall's patent description;
[0046]FIG. 4 is a circuit diagram showing a prior art regulator implemented with thermionic devices;
[0047]FIG. 5 is a general circuit diagram showing a NMOS depletion LDO regulator circuit in accordance with the present invention;
[0048]FIG. 6 is a circuit diagram showing a NMOS depletion LDO regulator circuit in accordance with the present invention;
[0049]FIG. 7 is a circuit diagram showing a NMOS depletion LDO regulator circuit in accordance with the present invention;
[0050]FIG. 8 is a circuit diagram showing a NMOS depletion LDO regulator circuit combining the error simplifier and voltage reference functions in accordance with the present invention; and
[0051]FIG. 9 is a circuit diagram showing a NMOS depletion LDO regulator circuit combining the voltage divider and voltage reference functions in accordance with the present invention.
[0053]FIG. 5 shows the most general embodiment for the low dropout voltage regulator 6 using the depletion MOS transistor MP1 as main pass element.
[0054] The linear regulator 6 comprises a voltage control circuit 7 to control the voltage at the gate of the transistor MD1 in order to regulate the voltage at the load.
[0055] Furthermore a current control circuit 8 controls the voltage applied to the gate of PMOS device MP1 in order to control the current to the load.
[0056] According to the embodiment of the present invention, the depletion pass transistor MD1 is configured as a follower to allow the gate voltage to regulate the voltage at its source. Its back gate could be shorted to the source, but in a more common embodiment is connected to the substrate of the device. Because it is a depletion mode device, MD1 requires a negative voltage at its gate relative to its source in order to be turned fully off.
[0057] The PMOS device MP1 in series with pass device MD1 allows the current to the load to be controlled even when the gate of MD1 cannot be driven negative with respect to the output, such as when the Vout terminal is at ground potential. MP1 can be controlled to be a constant current to act as a conventional current limit, or it can be made to be a function of the output voltage or other parameters, as for example, in a fold-back current limit that decreases the current limit value in the case of a short-circuited load.
[0059]FIG. 6 represents the general preferred embodiment for the low dropout voltage regulator using the depletion NMOS transistor as main pass element. FIG. 6 represents a more specific description of the system described in FIG. 5, with the current control block 8 and voltage control block 7 in FIG. 5 replaced with practical realizations.
[0060] The voltage control loop 7 of linear regulator 6 comprises a voltage reference circuit V2 having an output signal that connects to the non-inverting terminal of an operational amplifier A1, whose output controls the gate voltage of the main depletion pass transistor MD1 and whose inverting input connects to the feedback resistor divider implemented by R1 and R2.
[0061] The reference voltage V2 is most typically generated from a bandgap reference as is well known in the art. Other suitable references can also be derived, for example from a junction breakdown as with a zener diode, or from the difference between two dissimilar MOSFET or JFET thresholds. Although this reference is generally described as a constant voltage, this description does not preclude the use of a reference which has a functional value. For instance, a reference could be generated as a function of temperature to produce an output voltage Vout for regulator 6 which varies with temperature. Similarly, the reference voltage could be programmed, as with the output of a digital-to-analog converter, to make the regulator 6 programmable in output voltage.
[0062] The regulation is achieved by the operational amplifier A1 controlling the gate of MD1 in order to maintain the voltage at its two inputs at the same value. Therefore the output voltage will be regulated at the reference voltage multiplied by the resistor divider ratio.
[0063] The depletion NMOS transistor MD1 allows for a very low dropout voltage (difference between the input voltage and the output voltage) since its threshold is negative. With no substantive voltage between gate and source, as when V1 and Vout are at comparable levels, the NMOS MD1 will be turned fully on with a low resistance channel between drain and source.
[0064] Furthermore a large PMOS transistor MP1 is connected in series to the transistor MD1. Its gate is then connected to current control circuit 8 comprising a smaller PMOS MP2, current loop amplifier A2 and reference current I1. The transistors MP2 and MP1, together with the amplifier A2, form a current mirror with a gain determined by their channel width ratio, the channel length being preferably the same for both devices. The ratio of physical size is made preferably large, 1000 to 1 as shown in FIG. 6. Amplifier A2 works to force the current in MP2 to equal reference current I1 and for the voltage at the drain of MD1 and MD2 to be equal.
[0065] The operational amplifier A2 regulates the voltage at the drain of MD1 to be the same as the voltage at the drain of MP2. When the voltage at the drain of MD1 drops below the voltage at the drain of MP2 because the output current is approaching the current limit threshold, the operational amplifier A2 raises the voltage of the gate of MP1 and MP2 to control the current in the pass transistor MD1.
[0066] The current source I1 on the drain of MP2 is set to determine the output current limit as a multiple of the channel areas of MP1 and MP2. The generation of current sources is preferably independent of supply voltage and temperature, and is well known in the art of analog integrated circuits. The current source may also be made a function of input voltage, which can provide a constant power limiting, or can be made a function of temperature to increase the allowable dissipation when the die is cool, or as a function of the output voltage, to implement a fold-back limiting function. Other functional reasons for varying the current reference are foreseeable, and the general description of this current reference as a constant current source is not intended to limit such control of the current reference.
[0067] The regulator 6 will operate in one of two modes. When operating at a load current below the current limit, the output will be substantially controlled by the voltage control circuit 7. As the load current exceeds the current limit value, the output will be substantially controlled by the current control circuit 8.
[0068] When the current in the load is below the current limit, both PMOS MP1 and MP2 will be in the triode region. The effective resistance of the two devices will ratio as a function of their geometry, or 1000 to 1 as shown. When the current in MP1 and MD1 is substantially less than the current limit value, the drop across MP1 will be less than that across MP2. This will drive the inverting input of the amplifier A2 more positive than the non-inverting input, causing the output of amplifier A2 to swing low, further turning on both MP1 and MP2 until their gate voltages are substantially at ground potential. In this mode, MP1 is effectively turned on fully as a switch and MP1 plays no part in regulating the output.
[0069] When the current in the load increases to the value of current limit, the amplifier A2 actively regulates the current in MP1 as described above. Typically, as the current limit is reached, the output voltage will fall to a value below the ideal regulated voltage. The voltage at the inverting input of A1 decreases proportional to the output voltage. This drives the output of amplifier A1 positive and MD1 is turned fully on. The pass device MD1 becomes a fully enhanced switch in series with the current of the PMOS MP1 which effectively regulates the load.
[0071]FIG. 7 represents the first preferred embodiment for the low dropout voltage regulator using the depletion NMOS transistor MD1 as main pass element. This embodiment provides a more practical current control implementation than the general implementation of FIG. 6, and adds a logic input that can selectively enable or disable the operation of the regulator.
[0072] The linear regulator comprises a voltage reference circuit V2 having an output signal that connects to the non-inverting terminal of an operational amplifier A1, whose output controls the gate voltage of the main depletion pass transistor MD1 and whose inverting input connects to the feedback resistor divider implemented by R1 and R2. The voltage control loop so implemented is identical to that described above for FIG. 6.
[0073] Furthermore a large PMOS transistor MP1 is connected in series to the transistor MD1. Its gate is then connected to the gate of the PMOS transistor MP2 of the same type, but smaller channel size and to the drain of the PMOS transistor MP3 and to the output of current reference I1. The output of the current reference also connects to the drain of the PMOS transistor MP4. The gate of MP4 is connected to a terminal ENABLE which is used to selectively turn on or off the regulator. When ENABLE is substantially in the high state, then MP4 is off and the regulator works as previously described. When ENABLE is substantially low, the reference current from I1 is effectively shunted away from MP2 and MP3, such that MP1 is programmed for zero current and the regulator will produce no load current.
[0074] A second operational amplifier A2 has its inverting input connected to the drain of MP2, its non-inverting input connected to the drain of MD1 and its output to control the gate of the transistor MP3.
[0075] The amplifier A2 performs a function identical to that of A2 in FIG. 6, simultaneously forcing the current in MP2 to equal the reference current I1 and the drain voltage of MP2 to equal that of MP1. The inclusion of MP3 within this function simplifies stability of the loop by separating the control of these two simultaneous conditions. MP3 acts as a PMOS source follower, allowing the combination of amplifier A2 and PMOS MP3 to form a conventional unity gain follower forcing the drain voltage of MP2 to be substantially equal to the drain voltage of MP1, limited only to the input error on A2 as is well known in the design of operational amplifier circuits. Because the drain currents in MP2 and MP3 will be identical, any error between the value of current in MP2 relative to the magnitude of I1 will result in the difference in current flowing into the gates of MP1 and MP2. If MP2 is, for example, operating at a current slightly lower than the current reference I1, the excess current being sunk by I1 will flow from the gates of MP2 and MP1, lowering the voltage on the gates and therefore turning these devices on further. As the current in MP2 grows to equal that of I1, the gate current will reduce to zero and a stable condition will be reached.
[0076] A limitation of this current control circuitry compared to that of FIG. 5 is that the condition that drain voltages of MP1 and MP2 being held equal is maintained over a narrower range of voltages. As the voltage at the drain of MP1 falls below the voltage of the common gates of MP1 and MP2, amplifier A2 drives the gate of MP3 substantially to ground, turning on MP3 fully as a switch. The circuit thus formed will be recognized as a simple two transistor current mirror with the drain and gate of MP2 being effectively connected together through the low impedance of switch MP3. In this configuration, the circuit continues to operate as described above but the current in MP1 will have an error due to output impedance as is well known in simple current mirrors.
[0078]FIG. 8 displays another embodiment of the present invention for the low dropout voltage regulator using the depletion NMOS MD1 transistor as main pass element. This embodiment is similar to that of FIG. 7 but demonstrates that the functions of the error amplifier A1 and the reference voltage V2 of previous figures can be practically merged.
[0079] The linear regulator includes a voltage reference circuit 9 (of the type analogous to the Brokaw band-gap cell). The voltage reference appears at the gate of the NPN transistors Q1 and Q2, Q2 having its emitter area 10 times greater than the emitter area of Q1. The resistor R4 is connected to the emitter of Q2 and to the emitter of Q1 and the resistor R3 is connected between emitter of Q1 and ground. The PNP transistors Q3 and Q4 are connected in a current mirror configuration of conventional design to force Q1 and Q2 to operate at substantially equal current. The gate of the depletion NMOS pass transistor is connected to the collectors of Q1 and Q2.
[0080] Furthermore a large PMOS transistor MP1 and related current control circuitry is connected in series to the transistor MD1 and it operates as described for the case of the embodiment of FIG. 7.
[0081] Voltage regulation is achieved as the Brokaw cell band-gap circuit 10 controls the voltage at the gate of MD1, in order to maintain the voltage at the mid point of the resistor divider R1-R2 at the band-gap voltage (1.23V). Therefore the output voltage will be regulated at the reference voltage (typically the band-gap voltage) multiplied by the resistor divider ratio. As the voltage at the bases of Q1 and Q2 deviate from this preferred value, the collector currents in Q1 and Q2 become unbalanced. The collector current in Q4 is substantially equal to the collector currents of Q2 and Q3, and will therefore become unbalanced with respect to the collector current in Q1. This current imbalance creates a net current either charging or discharging the gate of MD1, which will change the voltage at Vout until the voltage at the bases of Q1 and Q2 regain their preferred value that will again balance their collector currents.
[0082] The transistor MP4 simply operates as a switch to disable the regulator guaranteeing zero output current as in the case of the embodiment shown in FIG. 7.
[0084]FIG. 9 shows one alternative embodiment for the low dropout voltage regulator using the depletion MOS transistor MD1 as main pass element. This embodiment is similar to that of FIG. 7 but demonstrates that the functions of the voltage reference V2 and the voltage divider R1 and R2 of previous figures can be practically merged. This implementation also shows a further improvement in the current control loop.
[0085] The linear regulator includes a Bandgap reference circuit 10 having two diodes D1 and D2, with D1 area ten times the area of D2 with their anode connected to ground. The cathode of D1 is further connected to the resistor R8, while the cathode of D2 is connected to the resistor R6 and to the non-inverting input of the operational amplifier A1 whose inverting input is connected to the resistors R8 and R7 and its output to the gate of the depletion NMOS pass transistor MD1.
[0086] Furthermore a large PMOS transistor MP1 is connected in series to the depletion pass transistor MD1 and it operates as described the embodiment of FIG. 7. Its gate is connected to the transistor MP2 of the same type, but smaller channel size and to the output of the operational amplifier A3.
[0087] A second operational amplifier A2 operates as described for the embodiment of FIG. 7.
[0088] A non-inverting amplifier A3 acts as a voltage buffer to generate a voltage shift of the voltage at the drain of MP3 to the gate of the transistors MP1 and MP2. This amplifier maintains its input voltage at a substantially low value such that the PMOS MP3 will not enter the triode region as the drain of MP1 drops in voltage when the regulator is in current limit.
[0089] The voltage reference is generated at the node that connects the resistor RS, R6 and R7. Resistors R6 and R7 are preferably made substantially equal. The regulation is achieved as amplifier A1 controls the gate of MD1 in order to maintain the voltage at its two inputs at the same value. The current in the two diodes D1 and D2 is substantially equal and the voltage across R8 is substantially the temperature dependent ΔVd that occurs when operating diodes at differing current densities. The voltage at Vout when the inputs of A1 are substantially equal is the sum of a diode voltage and of a voltage which is a scaled version of said ΔVd. The negative temperature coefficient of the diode voltages can be balanced against the positive temperature coefficient voltage imposed across the resistors. This balance occurs when the resistors are adjusted such that the total voltage is approximately 1.23V, the bandgap of silicon.
[0090] The series PMOS MP1 connected to the drain of MD1 operates as described for the embodiment of FIG. 7.
[0091] The transistor MP4 operates as described for the embodiment of FIG. 7.
[0092] Although the present invention has been described above with particularity, this was merely to teach one of ordinary skill in the art how to make and use the invention. Many additional modifications will fall within the scope of the invention. Thus, the scope of the invention is defined by the claims which immediately follow.
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Tech Helproom
It's free to register, to post a question or to start / join a discussion
ebook PDF files showing up on google when it was supposed to be encrypted
Likes # 0
Posted February 26, 2012 at 12:34PM
I am trying to sell ebooks on my website. The ebooks consist of questions and answers. People pay to download the answers. However, I have found that if I type the question into the google search it comes up with the PDF file that is in the root folder of my website, and people can just open it without paying anything. Why do you think this is?
Likes # 0
Posted February 26, 2012 at 1:51PM
Thanks. Yes, I am using Kindle.
I have not tried it on someone else's computer. I will try that.
I have now found the Robots section in my website builder and have disallowed the search engine spiders from indexing the pages which have the 'download now' button on them. But the PDF files are still coming up on my computer in google search.
Likes # 0
Posted February 26, 2012 at 3:06PM
I have now tried it on someone else's computer. The PDF files are still coming up on the google search and you can open them.
Likes # 0
Posted February 26, 2012 at 11:32PM
It might be better to send people a link when they pay for the answers, rather than linking to the files from a button.
Create a new directory in the root, and put the PDF files into it. Set up an auto-mailer that sends a link to the files once payment is received.
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subset: commoncrawl


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Jerry Williams
Follow Jerry all over social media.
Click the photo to get his Facebook page.
(scroll down for the feed
from the blog)
And then there's his Twitter:
Don't forget Instagram
Show Highlights
The quest for the perfect cup of coffee continues
Back in January, Rob Langer introduced us to what we've dubbed "Rob's Butter Java," coffee with a tablespoon of butter instead of cream. But not just any butter, this has to be butter from grass-fed cows.
Then last week Rob came back with another ingredient, organic coconut oil. This recipe is called "Langer Latte."
How many more coffee recipes can we come up with? Well, how about coffee with butter from coconut-fed cows? Not sure how easy it'll be to find that ingredient, but we'll keep experimenting.
read more
Jerry Williams: Haiku Master
One man's snack is Jerry's inspiration for Haiku!
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Social Media Milestones Within Reach
Granted, this may fall into the "shameless self promotion" category, but here goes. I'm within sight of a couple of milestones on social media. OK, these are not huge milestones when compared to other JOY FM staffers, but it is what it is.
On Twitter (which I haven't been very prolific on lately) I'm at almost at 200 followers.
You can help by "following" me on Twitter. Just click here.
I'm doing a little better on Facebook, where I'm closing in on 400 "likes."
You can help by "liking" my Facebook page here.
THANKS! I thinkk you'll enjoy the information and fun stuff I share through social media.
read more
Remembering Andrae Crouch
Andrae Crouch went home to be with the Lord Thursday, January 8, 2015. He was 72. He leaves behind an amazing legacy of service and musical innovation. Jerry shares some personal memories of Andrae on his blog.
Here's a video of one of Andrae's best known and most loved songs, with an all-star choir of Christian music greats, My Tribute (To God Be The Glory)
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This cardi is so cute for layering to/from yoga. It is slightly shorter in the front, longer in the back. There is a snap to secure the front sides together, although I'm not sure you would ever need to. The thumb holes are nice too.
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This Tuesday, March 24, 2015, the Drupal community lost Aaron Winborn who was diagnosed with ALS a few years ago. In honor of Aaron, the Drupal Association and Angie Bryon recently announced the Aaron Winborn Award. The announcement reads:
In honor of long-time Drupal contributor Aaron Winborn (see his recent Community Spotlight), whose battle withAmyotrophic lateral sclerosis (ALS) (also referred to as Lou Gehrig's Disease) is coming to an end later today, theCommunity Working Group, with the support of the Drupal Association, would like to announce the establishment of the Aaron Winborn Award.
This will be an annual award recognizing an individual who demonstrates personal integrity, kindness, and above-and-beyond commitment to the Drupal community. It will include a scholarship and stipend to attend DrupalCon and recognition in a plenary session at the event. Part of the award will also be donated to the special needs trust to support Aaron's family on an annual basis.
Thanks to Hans Riemenschneider for the suggestion, and the Drupal Association executive board for approving this idea and budget so quickly. We feel this award is a fitting honor to someone who gave so much to Drupal both on a technical and personal level.
Thank you so much to Aaron for sharing your personal journey with all of us. It's been a long journey, and a difficult one. You and your family are all in our thoughts.
Jahia, a leading open source User Experience Platform (UXP) vendor, announced today that Piyush Patel, joins Jahia CEO Elie Auvray and COO Emmanuel Garcin in the Executive Leadership team of the company as Executive Vice President of Strategic Alliances & General Manager Americas.
London, 10 March 2015 - Creating everything as a service (XaaS) and delivering innovation is set to be the major driver of IT development this year as XaaS becomes the preferred consumption model for enterprises. In its conference sessions, the European Software and Solutions Summit (incorporating the eighth European ISV Convention) will feature speakers from Gartner, HP, Interoute, Oracle, Salesforce.com and Verizon delivering their visions of how this changes how users buy IT, who buys it and how it is paid for.
More and more, businesses expect IT to enable their staff to work wherever they are located and via whatever device is most appropriate, says Duncan Campbell Senior Director, EG Alliances Marketing and Business Development Hewlett-Packard Company who is one of the presenters. "They want to have technology on-demand that can scale and is secure, PLUS pay only for what they consume. They want IT to innovate with the line of business to enable them to transform their processes and the user experience. Those IT departments that do not engage with the business will not be relevant, and their budgets will be impacted."
Scott Liewehr emailed me today announcing that Digital Clarity Group and Constellation Research, Inc. are forming a strategic alliance to better serve their clients. Scott is CEO of Digital Clarity Group and it's no secret CMS Report is an admirer of both him and his company. Yet Scott's email still surprised me. Why would DCG, rising among the ranks among analyst firms, strike an alliance with Constellation?
The press release that accompanied Scott's email gives reasons for the strategic alliance, but I'll add one observation of my own. Both DCG and Constellation likely recognize they share the same understanding of how legacy research firms are failing their clients and believe a fresh approach to handling "digital disruption" is necessary. An understanding that can easily translates to the buyer seeking out this kind of research that two heads are better than one.
These days, content is the backbone of online business, impacting customer decision-making before direct contact is made with sales or through marketing automation. As channels proliferate, Enterprises need to ensure that they deliver personalized content that resonates with their diverse audiences, in any context and on any device.
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To ensure that their content is helping reach business goals, enterprises need a smart, third-platform Web Content Management System that uses data and metrics to continuously optimize their content's performance.
TERMINALFOUR, the digital engagement and web content management platform for higher education, today announced its plans to expand its North American operations and recruit 15 new positions in its Boston headquarters. The expansion is being accelerated due to the successful launch of TERMINALFOUR's latest software release which has resulted in TERMINALFOUR winning 7 new significant higher education clients.
This is the third year for the Now What? Conference which is held in one of the fastest growing modern communities in the region, Sioux Falls, SD. If you're within a half day's drive from this great city, I encourage you to register for the conference. The conference and various workshops will be held on April 29 - 30, 2015. Created by Blend Interactive, the conference will be bringing together web and marketing professionals from across North America. Something that rarely happens in our region, you have an opportunity to hear from today's content management leaders as they come together and cover post-launch web maintenance, web analytics, content strategy, and talk shop with colleagues and speakers.
The digital learning content management business solution provider, eXact learning solutions, has revealed a number of refinements to its learning content management system (LCMS) and its constituent suite of products.
The Board of Directors of Paragon Development Systems (PDS, Inc.), a technology solutions and services company, has named Asif Naseem as the company's new President and CEO.
Propeople, a full-service agency for the digital age, today announced that it is expanding its Global Executive Team with the appointment of two new positions - Gus Murray as Global Chief Digital Strategy Officer, and Troels Feodor Nielsen as Global Chief Marketing Officer. Both Gus and Troels will help to steer Propeople's strategic initiatives in current and future markets.


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Page 1
JOURNAL OF VIROLOGY, Dec. 2008, p. 12569–12573
Copyright © 2008, American Society for Microbiology. All Rights Reserved.
Vol. 82, No. 24
Role of Ebola Virus VP30 in Transcription Reinitiation?
Miguel J. Martínez,1,2† Nadine Biedenkopf,3,4† Valentina Volchkova,1,2,5,6Bettina Hartlieb,3,4
Nathalie Alazard-Dany,1,2‡ Olivier Reynard,1,2Stephan Becker,3,4* and Viktor Volchkov1,2,5,6*
INSERM, U758, Filovirus Laboratory, 21 Av. Tony Garnier, Lyon F-69007, France1; IFR 128, Lyon F-69007, France2; Robert-Koch-Institut,
ZBS5 Berlin, Germany3; Institut fu ¨r Virologie, Philipps Universita ¨t Marburg, Marburg, Germany4; Universite ´ de Lyon,
Lyon F-69007, France5; and Universite ´ de Lyon 1, Villeurbanne F-69622, France6
Received 3 July 2008/Accepted 19 September 2008
VP30 is a phosphoprotein essential for the initiation of Ebola virus transcription. In this work, we have
studied the effect of mutations in VP30 phosphorylation sites on the ebolavirus replication cycle by using a
reverse genetics system. We demonstrate that VP30 is involved in reinitiation of gene transcription and that
this activity is affected by mutations at the phosphorylation sites.
Ebola virus (EBOV) causes a severe hemorrhagic fever syn-
drome, characterized by high mortality rates, in humans and
nonhuman primates (7, 13, 14). The EBOV replication cycle
takes place in the cytoplasm of infected cells, where inclusion
bodies are formed (4). The inclusions contain viral ribonucleo-
protein (RNP) complexes consisting of viral RNA and four
nucleocapsid proteins: the nucleoprotein (NP), VP35, the
polymerase (L), and VP30 (15, 16). Using an EBOV mini-
genome system, it was demonstrated that NP, VP35, and L are
sufficient for RNA replication, whereas the addition of VP30 is
required for transcription initiation (12). Recently, it has been
shown that VP30 provided in trans supports transcription of
the minigenome delivered by infectious virus-like particles
(VLPs) and replication of a recombinant EBOV lacking the
VP30 gene (5). VP30 is phosphorylated at two serine clusters
(amino acids 29 to 31 and 42 to 46), each containing three
serine residues (Fig. 1A). Previously, it has been shown that
mutants of VP30 with both serine clusters replaced by alanine
residues supported transcription of an EBOV minigenome but
did not accumulate in the NP-induced inclusions. In contrast,
when all serine residues at the phosphorylation sites were
replaced by aspartate residues, VP30 was unable to support
transcription (10). While a Ser3Ala substitution mimics non-
phosphorylated serine, a Ser3Asp substitution mimics con-
stantly phosphorylated serine.
In this study, we investigated the effect of mutations simu-
lating a constantly phosphorylated state of VP30 during a
Zaire EBOV infection (Fig. 1A). VP30 mutants with nonphos-
phorylated and highly phosphorylated states are represented
by VP30-AA and VP30-DD, respectively. Two other con-
structs, VP30-AD and VP30-DA, have one of the two serine
clusters replaced by an alanine cluster and the other by an
aspartate cluster.
In an attempt to generate recombinant viruses containing
the designated mutations, we used a reverse genetics system
for EBOV (17). Only the wild-type EBOV genome was res-
cued into infectious virus (Fig. 1B). Failure to recover a virus
containing VP30-DD is likely explained by the lack of tran-
scription initiation activity of VP30-DD (10). To assess
whether the ability of VP30 to support viral transcription was
altered with the other three mutants, we quantified their ac-
tivities in an EBOV-specific minigenome system (8) (Fig. 2A
and B). VP30-AA was 30% more active than the VP30 wild
type (VP30-WT) in supporting EBOV transcription, and both
VP30-AD and VP30-DA supported transcription of the re-
porter gene in the same range as VP30-WT. Thus, the exper-
iments with minigenomes did not provide a plausible explana-
tion for the failure to recover recombinant viruses containing
the AA, AD, or DA mutations.
In addition to transcription initiation activity, VP30 is a
structural protein (3, 6, 14). Since it has been shown that
mutations at the phosphorylation site alter VP30's association
with NP-induced inclusions (10), we presumed that mutations
at the phosphorylation site would impact the incorporation of
VP30 into viral particles and thus affect the efficiency of virus
recovery. To assess the ability of VP30 mutants to support viral
morphogenesis, we used an EBOV-specific VLP system (8).
We found that all mutants of VP30 were incorporated into
VLPs and were protease protected (Fig. 2C). This indicated
that VP30 mutants were localized inside the particles. To con-
firm these results, we tested whether the mutants of VP30 were
also incorporated into authentic EBOV. Subconfluent 75-cm2
flasks of Vero cells were transfected with 8 ?g of plasmids
encoding myc-tagged mutants of VP30 and were then infected
with EBOV at a multiplicity of infection (MOI) of 1 at 6 hours
posttransfection. Virions released into the culture medium
during 24 h of infection were collected by ultracentrifugation
through a 20% sucrose cushion and then analyzed by Western
blotting using anti-VP24 and anti-myc antibodies. Both
VP30-AA and VP30-DD mutants were found to be associated
with viral particles (Fig. 2D). These data are in agreement with
a recent study (6) showing that recruitment of VP30 into VLPs
is dependent on the C-terminal part of the protein, while our
* Corresponding author. Mailing address for Viktor Volchkov:
INSERM, U758, Filovirus Laboratory, 21 Av. Tony Garnier, 69365
Lyon, Cedex 07, France. Phone: 33 437 282450. Fax: 33 437 282459.
E-mail: firstname.lastname@example.com. Mailing address for Stephan
Becker: Institut fu ¨r Virologie, Philipps-Universita ¨t Marburg, 35037
Marburg, Germany. Phone: 49 6421 2866253. Fax: 49 6421 2868899.
E-mail: firstname.lastname@example.com.
‡ Present address: INSERM, U758, Ecole Normale Supe ´rieure, 46
alle ´e d'Italie, 69364 Lyon, Cedex 07, France.
† These authors contributed equally to this work.
?Published ahead of print on 1 October 2008.
Page 2
mutations affected the N terminus. Moreover, our results sug-
gest that the intracellular colocalization of VP30 and NP de-
scribed in an earlier publication (10) does not reflect the ability
of VP30 to be incorporated into budding VLPs. It should be
mentioned that the smaller amount of VP30-DD in pelleted
virions compared with that of VP30-WT and VP30-AA was not
expected based on our results with VLPs. However, this is
explained by the negative effect of VP30-DD on viral replica-
tion. Since only approximately 10 to 15% of cells were trans-
fected and expressed mutated VP30, this effect is not seen at
the level of the VP24 expressed in all virus-infected cells. To
confirm the effect of VP30-DD on viral replication, 293T cells,
which have much higher transfection efficiency, were used
(Fig. 2E).
Taken together, our results showed that VP30-WT, VP30-
AA, VP30-AD, and VP30-DA were similar in terms of their
association with the nucleocapsid and transcription of EBOV
minigenomes, but they differed in their capacities to rescue
recombinant viruses. One possible explanation of their differ-
ences is that in the minigenome system, a monocistronic tem-
plate is transcribed, whereas rescue of full-length viral RNA
needs transcription of seven individual genes in a consecutive
manner. At every gene end, transcription stops and is reiniti-
ated at the start of the following gene (11). We therefore
analyzed the capacities of the VP30 mutants to reinitiate tran-
scription. The plasmid encoding VP30-WT was replaced by
others encoding VP30-DD, VP30-AA, VP30-AD, or VP30-DA
in the full-length rescue system in combination with the wild-
type EBOV genome (Fig. 3). If the mutants of VP30 are able
to support transcription of all viral genes, VP30, which is en-
coded by the fifth gene of the viral genome, will be synthesized
and will compensate for putative missing functions of the mu-
tant VP30. Here, we demonstrate that VP30-DD, VP30-AD,
and VP30-DA do not support the recovery of the wild-type
FIG. 1. (A) Nomenclature of VP30 mutants. Two clusters of serine residues (S29, S30, and S31; S42, S44, and S46) constitute the protein
phosphorylation sites. Several cellular protein kinases (e.g., PKA, CKII, and cAMKII) are predicted to recognize VP30 phosphorylation sites.
Amino acid substitutions and the nomenclature of the constructed mutants are indicated. (B) Recovery of recombinant viruses. (Left) Schematic
representation of the reverse genetics system. Mutations in the VP30 gene are introduced in a plasmid carrying a full-length cDNA of Zaire EBOV.
The recovery of the recombinant virus is then supported by four plasmids encoding wild-type nucleocapsid proteins (NP, VP35, VP30, and L).
(Right) Results of the recovery experiment. Successful recovery of the virus is indicated by a plus. Lack of virus recovery is indicated by a minus.
Page 3
EBOV. In contrast, VP30-AA supported the recovery of the
wild-type EBOV genome, indicating that the nonphosphory-
lated form of VP30 activates transcription of all viral genes,
including the gene encoding VP30. Since VP30-AD and
VP30-DA showed an activity comparable to that of VP30-AA
in supporting transcription of the reporter gene, their failure to
recover the wild-type virus pointed to a difference in support-
ing transcription of downstream genes.
Next, we assessed the abilities of VP30-AD and VP30-DA to
support transcription reinitiation. We generated a plasmid car-
rying a full-length cDNA of EBOV containing the green fluo-
rescent protein (GFP) gene as an additional transcription unit
between the genes encoding VP35 and VP40 (the second and
third genes). When VP30-AA was used to rescue this virus,
GFP-expressing cells appeared 2 days posttransfection and
gradually increased in number, corresponding to the spread of
the recombinant virus (Fig. 4A). In contrast, no GFP-express-
ing cells were observed in the presence of VP30-AD or VP30-
DA, suggesting that these mutants did not support transcrip-
tion of the GFP gene. These results strongly support the idea
FIG. 2. Influence of permanent charges at the phosphorylation sites of VP30 on viral transcription and incorporation of VP30 into virus
particles. (A) Schematic representation of the minigenome and VLP assay. HEK293 cells were transfected with plasmids encoding all viral
proteins, the T7 polymerase, and the EBOV-specific minigenome (MG) containing a Renilla luciferase reporter gene. Additionally, the vector
pGL4 carrying the firefly luciferase reporter gene was transfected and used for normalization of the results. Nucleocapsid proteins provided in trans
direct transcription/replication of the EBOV minigenome, which is subsequently encapsidated into VLPs.
minigenome containing the luciferase reporter. (B) Transcription activation by VP30-WT and VP30 mutants. At 72 h posttransfection, cells were
harvested and analyzed for reporter activity using the Dual-Luciferase assay (Promega). Renilla luciferase activity (R-Luc), reflecting minigenome
replication and transcription, was normalized by firefly luciferase activity. Transcription activation by VP30-WT was set to 100%. Replacement of
VP30-WT by the respective mutants is indicated. Rel., relative. (C) Incorporation of flag-tagged VP30 into VLPs. VLPs from culture supernatants
collected at 72 h posttransfection were purified by ultracentrifugation through a 20% sucrose cushion. Incorporation of VP30-WT and mutants of
VP30 was analyzed by a protease protection assay. Samples were treated with proteinase K in the presence or absence of Triton X-100, separated
by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and then detected by Western blotting using antiflag antibodies. Proteins inside
VLPs were protected from proteinase K digestion and are visible in the middle columns. (D) Incorporation of myc-tagged VP30 proteins in EBOV
particles. Vero E6 cells were transfected with plasmids encoding myc-tagged VP30 (VP30-WT [WT], VP30-AA [AA], or VP30-DD [DD]) and then
infected with EBOV at an MOI of 1 PFU/cell. Virus particles from culture supernatants were purified by ultracentrifugation through a 20% sucrose
cushion. NT, nontransfected cells. (E) Expression of VP30-DD negatively affects EBOV replication. 293T cells were transfected with plasmids
expressing either VP30-WT or VP30-DD and then infected with EBOV at an MOI of 1 PFU per cell. Samples of culture supernatants (SN) and
lysed cells were analyzed by Western blotting using anti-VP24, anti-NP, and antiactin antibodies.
, transcription initiation of the
Page 4
that the AD and DA substitutions in VP30 affected the ability
of the polymerase complex to reinitiate transcription of viral
genes downstream of the second gene.
To confirm that VP30-AA was able to reinitiate transcrip-
tion without the expression of VP30-WT, we mutated the full-
length cDNA of EBOV containing the GFP gene by introduc-
ing the AA mutation in the VP30 gene and employed this
construct in the rescue system by expressing the different mu-
tants of VP30 in trans. Individual cells expressing GFP were
observed when VP30-AA was employed, whereas no expres-
sion of GFP was detected when VP30-AD or VP30-DA was
employed (Fig. 4B). This result confirmed that VP30-AA was
able to support viral transcription of the first gene as well as
consecutive genes. In contrast, the mutants VP30-AD and
VP30-DA, while capable of initiating transcription of the first
gene, were unable to reinitiate transcription of downstream
genes. Besides, these data suggest that a cluster of negative
charges mimicking phosphorylation could inhibit VP30's tran-
scription reinitiation function.
Our experiments demonstrate that the serine clusters at the
N terminus of VP30 represent a critical region for the activity
of the protein. A combination of alanine and aspartate resi-
dues (VP30-AD and VP30-DA) resulted in a VP30 protein
that was able to support transcription of an EBOV-specific
monocistronic minigenome but unable to reinitiate transcrip-
tion in the full-length genome. Permanently uncharged amino
acids at the phosphorylated region (VP30-AA) resulted in
VP30 molecules that were able to activate transcription of a
minigenome and all the genes of the EBOV genome in a
consecutive manner. However, a recombinant EBOV encoding
VP30-AA could not be generated, indicating that this mutant
lacks another function needed to produce fully infectious viri-
ons. The step at which phosphorylated VP30 is essential in the
viral life cycle is currently unknown. Since VP30-DD blocks
EBOV transcription, the phosphorylation of VP30 might play
a role in switching off the transcription favoring replication of
the viral RNA. In this regard, the permanently nonphosphor-
ylated VP30-AA would impair the regulation of the transcrip-
tion/replication processes. In addition, since a phosphorylated
form of VP30 is present in EBOV virions (3), phosphorylated
VP30 may be necessary for a very early step in the viral life
cycle. Our results thus suggest that the dynamic phosphoryla-
tion of VP30 is essential for the function of the protein in
EBOV infection.
It is presumed that transcription in viruses from the order
Mononegavirales follows a sequential stop-start mechanism in
which transcription of downstream genes is dependent on ter-
mination of the synthesis of the upstream gene. While the
majority of studies have focused on the influence of cis-acting
sequences in transcription regulation (1, 2, 19), in our work, we
provide experimental evidence of the participation of a struc-
tural protein other than the viral polymerase in transcription
reinitiation. The function of VP30 in reinitiation of transcrip-
tion expands on results from a previous report showing that a
specific stem-loop structure at the beginning of the NP gene is
involved in VP30-dependent transcription and that only tran-
scription initiation of the NP gene requires VP30 (18). In the
artificial bicistronic EBOV minigenome, transcription of the
second gene did not require the activity of VP30. Using a
full-length EBOV genome, we obtained results that partially
FIG. 3. Rescue of wild-type EBOV using mutants of VP30. (Left)
Schematic representation of the experimental setup. The recovery of a
wild-type recombinant EBOV is supported by plasmids expressing the
nucleocapsid proteins NP, VP35, and L together with mutants of
VP30. (Right) Results of the recovery experiment. Successful recovery
of the virus is indicated by a plus. Lack of virus recovery is indicated by
a minus.
FIG. 4. Recovery of recombinant EBOV expressing GFP. The GFP gene was introduced into the full-length cDNA of either wild-type EBOV
(A) or EBOV/VP30-AA (B). Recombinant virus recovery was performed in BHK T7 cells using standard protocols and different VP30-expressing
plasmids as indicated. The presence of groups of green cells indicated recovery and replication of the recombinant viruses. The appearance of
single green cells indicates the ability of mutated VP30 to support both transcription initiation and reinitiation but with subsequent failure of virus
replication. , transcription initiation;, transcription reinitiation.
Page 5
contradict this earlier report. This, however, only highlights the
need to complement minigenome studies with the analysis of
full-length genome expression systems. Recently, EBOV VP30
was shown to be an RNA-binding protein (9). Notably, VP30
binds to the sequence in the NP gene's stem-loop structure that
includes the transcriptional start signal and its complementary
sequence. Sequences complementary to the respective tran-
scription start signal are present in close proximity to the start
signal of all EBOV genes, suggesting that the stem-loop struc-
tures also play a role in the transcription of other genes (11).
Interestingly, analysis of these structures using the mfold pro-
gram (20) showed that their energetic stability (?G) is in-
creased in a 3?35? order (NP, ?7.40; VP35, ?8.6; VP40,
Given that VP30 is involved in transcription reinitiation, it is
reasonable to speculate that there is a gradient along the viral
genome that controls the efficiency of transcription, especially
at the 5? genome end.
In conclusion, our data indicate, for the first time, that ac-
tivity of VP30 is required for reinitiation of gene transcription
and that mutations at the phosphorylation sites of VP30 affect
this activity. While the mechanism of VP30 action is not yet
clearly understood, our results suggest that VP30 could be an
important target for antiviral therapy.
All experiments involving live EBOV were carried out in the
INSERM BSL-4 laboratory Jean Merieux in Lyon, France. We thank
the biosafety team members for their support and assistance in con-
ducting experiments and Ulla Thiesen for expert technical assistance.
We are also grateful to Robin Buckland for his helpful comments on
the manuscript.
This work was supported by INSERM, the French Ministe `re de la
Recherche (grant 04G537), the Agence National de la Recherche
(ANR), the National Institutes of Health (grant AI059536), and the
Deutsche Forschungsgemeinschaft (SFB 593 to V.V. and SFB 535 to
S.B.). M.J.M. was partially supported by the Sociedad Espan ˜ola de
Enfermedades Infecciosas y Microbiologia Clinica (SEIMC). N.B. is
supported by the Ernst Schering Foundation.
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netics and manipulation of viral genomes. Annu. Rev. Genet. 32:123–162.
2. Conzelmann, K. K. 2004. Reverse genetics of mononegavirales. Curr. Top.
Microbiol. Immunol. 283:1–41.
3. Elliott, L. H., M. P. Kiley, and J. B. McCormick. 1985. Descriptive analysis
of Ebola virus proteins. Virology 147:169–176.
4. Geisbert, T. W., and P. B. Jahrling. 1995. Differentiation of filoviruses by
electron microscopy. Virus Res. 39:129–150.
5. Halfmann, P., J. H. Kim, H. Ebihara, T. Noda, G. Neumann, H. Feldmann,
and Y. Kawaoka. 2008. Generation of biologically contained Ebola viruses.
Proc. Natl. Acad. Sci. USA 105:1129–1133.
6. Hartlieb, B., T. Muziol, W. Weissenhorn, and S. Becker. 2007. Crystal struc-
ture of the C-terminal domain of Ebola virus VP30 reveals a role in tran-
scription and nucleocapsid association. Proc. Natl. Acad. Sci. USA 104:624–
7. Hoenen, T., A. Groseth, D. Falzarano, and H. Feldmann. 2006. Ebola virus:
unravelling pathogenesis to combat a deadly disease. Trends Mol. Med.
8. Hoenen, T., A. Groseth, L. Kolesnikova, S. Theriault, H. Ebihara, B. Hart-
lieb, S. Bamberg, H. Feldmann, U. Stro ¨her, and S. Becker. 2006. Infection of
naïve target cells with virus-like particles: implications for the function of
ebola virus VP24. J. Virol. 80:7260–7264.
9. John, S. P., T. Wang, S. Steffen, S. Longhi, C. S. Schmaljohn, and C. B.
Jonsson. 2007. Ebola virus VP30 is an RNA binding protein. J. Virol.
10. Modrof, J., E. Muhlberger, H. D. Klenk, and S. Becker. 2002. Phosphoryla-
tion of VP30 impairs ebola virus transcription. J. Biol. Chem. 277:33099–
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Becker. 1996. Termini of all mRNA species of Marburg virus: sequence and
secondary structure. Virology 223:376–380.
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Comparison of the transcription and replication strategies of Marburg virus
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13. Peters, C. J., and J. W. LeDuc. 1999. An introduction to Ebola: the virus and
the disease. J. Infect. Dis. 179(Suppl. 1):ix–xvi.
14. Sanchez, A., T. W. Geisbert, and H. Feldmann. 2007. Filoviridae: Marburg
and Ebola viruses. In D. M. Knipe, P. M. Howley, and D. E. Griffin (ed.),
Fields virology, 5th ed. Wolters Kluwer/Lippincott Williams & Wilkins, Hag-
erstown, MD.
15. Sanchez, A., M. P. Kiley, B. P. Holloway, and D. D. Auperin. 1993. Sequence
analysis of the Ebola virus genome: organization, genetic elements, and
comparison with the genome of Marburg virus. Virus Res. 29:215–240.
16. Volchkov, V. E., V. A. Volchkova, A. A. Chepurnov, V. M. Blinov, O. Dolnik,
S. V. Netesov, and H. Feldmann. 1999. Characterization of the L gene and 5?
trailer region of Ebola virus. J. Gen. Virol. 80:355–362.
17. Volchkov, V. E., V. A. Volchkova, E. Muhlberger, L. V. Kolesnikova, M.
Weik, O. Dolnik, and H. D. Klenk. 2001. Recovery of infectious Ebola virus
from complementary DNA: RNA editing of the GP gene and viral cytotox-
icity. Science 291:1965–1969.
18. Weik, M., J. Modrof, H.-D. Klenk, S. Becker, and E. Mu ¨hlberger. 2002.
Ebola virus VP30-mediated transcription is regulated by RNA secondary
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Steven Smith TeamakerEdit Company
Popular Teas from Steven Smith TeamakerSee All 41 Teas
Recent Tasting Notes
Had been wanting to try this one for a while, now that I've tried it I can't remember quite why I wanted it so badly. It's definitely one of the weirdest teas I've ever tried! I really don't know what to think. 0_0
It's pine-y, kinda peppery, ever so slightly minty, and has some weird other spices going on that I can't pick out. It's seriously so weird. Might be the weirdest one yet!
It's not a horrible weird, but it's not an amazing weird either. I feel like it's the kind of weird that my taste buds will just like start craving randomly sometime for no apparently reason. It's a weird combo of flavors. But part of it kinda reminds me of a certain food. What food is that, you might ask? I don't know. xD But it really does remind me of something.
So yeah, not my kind of tea really. But thank you soo much for letting me try it, Rosehips!! :D :D
Flavors: Mint, Pepper, Pine, Spices, Wood
Basically the Lord Bergamot minus the bergamot. Honestly, not a favorite of mine. The quality is there though. I can totally imagine this being a go-to-tea for others. I still love their sachets, they work wonderfully and are so convenient.
Licorice and Peppermint.
I never thought that those two would mix.
When Rosehips sent me a sample of it, and I smelled the tea bags, I still wasn't sure. But man was I wrong!
Those two flavors together made for an excellent cup! I can't even explain how amazing it was! I wouldn't consider myself a huge fan of either of those flavors alone, but I haven't had many teas with those flavors in them. I really never could imagine they would mix so well.
PLUS! It's a chamomile!
I desperately wish that I had tried this before all of this tea was sold out. Now I have to wait a whole 'nother year to get more of this tea! I'm so sad! Thankfully Rosehips was so generous and gave me three tea bags. I will savor the two I have left!
I seriously had such a horrible day yesterday, and an even worse night. This tea was seriously the best part of my entire day last night. It helped me relax and actually smile and feel a little happier inside.
Ahh! I'm just so happy to have gotten to try this! And it really did cheer me up some last night. And writing this makes me smile because I'm really happy that I still have two more tea bags left! I should save them for the really bad days so they will help me feel more relaxed and a bit happier! (: I can't wait till Next Christmas to get more!
Thank you, Rosehips!! I'm so excited and happy about this tea! :D
Flavors: Licorice, Peppermint, Smooth, Sweet
Mmm, I know what you mean about liquorice and peppermint! It sounds really odd, but they work so well together as a tea! I've never tried this particular one, but the Teapigs one is ace :-)
Sample Sipdown!
Had the last of this tea last night. Not bad-all three times I had it it made for enjoyable and relaxing cups. Always happened to be when I needed it the most! I don't think I'd buy this tea in particular, but it was pretty nice. Thanks again for the generous samples, Rosehips!
I really really needed something to calm me down after this morning when I had to go to the dr to have a horrible test done. My body hurts like hell and I can barely swallow. Seriously have been such a mess today.
But this tea.
This tea totally calmed me down when I got home. Hurt tons to swallow, but it was definitely worth it. Helped me to be able to sleep for most of the day. xD Which was good for me because this monitor in my throat is killing me. xD
Also I really do think that the blend of lemon and chamomile is pretty nice in this tea. One tea bag left. :O
Thanks you guys. I'm sure that if the dr lets me come back and get this thing out of my throat himself it will be better. But kinda doubting he will. But fingers crossed! >
Got a generous sample of this tea from Rosehips! I really did need a relaxing tea, and this one totally worked! ^^
I love how much chamomile relaxes my muscles, it's like I feel it between the sips. It's the perfect kind of tea for me. It inspires me to try all of the chamomiles!
I wasn't sure if I'd like this one when I looked it up on Steepster, but I tried it anyway.
It's definitely interesting.
The main flavors are lemon and chamomile. They taste almost the same since chamomile is generally pretty sour tasting. Kinda a weird combo since it was a little hard to tell them apart, but I look forward to trying the other two tea bags of this one. It's interesting. Not something I'd go out and buy, but it really does make for a pleasant cup when you want to relax.
Thanks for the samples, Rosehips!
Flavors: Lemon, Sour
Like this tea quite a lot. Really my first taste of a bergamot but it was fantastic. I'm always a fan of Assam teas which is the base of this tea (with Ceylon). Just fantastic in my opinion. Impressive especially for a bagged tea (though they call them "sachets" which I suppose is accurate as they've got plenty of expansion room).
Flavors: Bergamot, Earth
Tea of the morning.
I was very interested. A rich blend of black teas, and a hint of pine? What would that be like?
I'm pleased to let you know it was not at all like a pine tree. I was afraid it might taste like a Christmas tree lot, but the pine note is lovely, subtle, and well blended with the black teas.
Good. I'm quite glad.
My collection of sachets are starting to get disturbingly low. That's going to be a bit of a problem, as I really like this, and its a seasonal tea!
It would be too simple to say that this is a mint or peppermint tea. Its so much more than that, full of a wonderful layer of lemon, and then a hint of licorice. The most sophisticated mint tea I've ever owned.
There is a very modern sweetness too this. Not an old fashioned, sugary sweetness, but something crisp and cool, and a little unexpected. It makes it much more than a mint tea, with the flavors running around and making themselves hard to identify.
I have a headache, and ate too much dinner, but this tea has helped take the edge off. Good!
Backlogged from the other day.
I brought this and a few other mint teas of mine over to my friends house, for a tea and knitting date. It was good! I'm getting so much progress done on the very basic scarf that I am learning to knit.
She picked this one, and so I had another chance to mull over my thoughts.
Still like it! Complex and layered. I think it will be great if you are not feeling well, which may sound clinical, but I think is a high bit of praise. You don't always want something this good when you don't feel well.
My friend was a little more meh about it, but thats alright!
So, I was starting to feel slight pangs about the amount of mint and peppermint based teas that I ordered from the Black Friday sales. Had I ordered too many? Would I get sick of similar flavors?
I was feeling slightly worried about this one, as I've jut discovered a love of the ROT Peppermint Bark. Would this taste the same?
I should not have worried. This is worlds different.
The smell was very light and sweet, and, when sipped, proved to be the most complex mint based tea I've ever had. Is that cinnamon in it? Orange? What were those other flavors? Whatever they were, they were fantastic. I will need many more cups of this fascinating blend to really pin it down.
Sophisticated may not always be the word used about peppermint tea, but it can be used without hesitation of Silent Night.
Thanks to boychik for this sample. I ended up steeping it for 3 minutes at 205 f and that was definitely overkill because I got really intense tobacco notes that made it difficult for me to drink this without milk. I also noticed spicy pepper-y notes and a bit of the sweet potato type of flavor that seems characteristic of yunnans.
I tried a 2nd steep for 90 seconds and now it seems like it doesn't have enough flavor… it could be I steeped all the flavor out of it at the 3 minute mark. :) Anyway the 2nd steep isn't bad but I'm still getting tobacco notes with some caramel. Tobacco isn't really my favorite flavor in tea.
this doesn't seem to be my favorite yunnan to date but it isn't terrible. If I had more tea I feel like I could hit a sweet spot with the brewing parameters. I wish I had more to play around with but probably not worth placing an order.
Very sweet, with a hint of raisin and lots of caramel. I can see why this is a favorite for so many on here, but, weirdly, I prefer my black teas on the harsher side—this Assam actually reminds me of some of the popular Chinese blacks I haven't cared for on grounds of having too much in the way of sweet dessert notes going on, though I can't think which ones exactly. Thanks for the sample, Cameron B.! I'm happy to be able to knock this off my wish list, since it's a tea I'd been curious about for a while.
This was a bonus sample from scribbles. Mine is in sachet form. The leaves inside are black and somewhat thin, and surprisingly not very broken. Dry scent is sweet and malty with hay and molasses notes. I let my sachet steep in 8 ounces of 200 degree water for 3 minutes. The string escaped into the cup, harumph!
This one is pretty "meh" in my opinion… The main flavor of it is generic black tea, and I don't know how else to describe that. It's somewhat malty and quite astringent, especially as it cools. There is a bottom note of molasses-like richness, and a touch of dried apricot-like fruitiness, and these help to balance it a bit. Still not very impressive for me.
Flavors: Apricot, Astringent, Malt, Molasses, Tea
This is going to be a lazy note for a lazy morning… Boychik sent me this sample, thanks dear. The leaves are fairly broken and dark with only a few scant golden pieces. I did my general black tea steep.
Meh, this one is just okay for me. It's fairly generic with just malt and some bread notes, along with a fair amount of astringency, which I do not enjoy. Maybe just a touch of honey sweetness on top. Overall, it's drinkable, but not something I would reach for.
Flavors: Astringent, Baked Bread, Honey, Malt
Thought I'd have this one again since they have a free shipping deal again. Had to see if I needed more. :P This really is a wonderful assam. I really do enjoy it. But I don't think I need more than 2oz for now. Maybe once I sip these 2oz down I'll be ready to buy more!
Still a very strong, malty assam. Enjoying this one!!
If you haven't had it yet and like malty teas-you should definitely take the free shipping offer up now and get some! It's not super expensive too!
Flavors: Malt
Decided to order this one since Steven Smith Teamaker is having a small sale, and since i'm working again I have more money! Glad I took a chance and got this one.
It's delicious! :O
Rich. Strong. Malty. Fruity. Honey.
A lovely tea!
And guess what! NO WOODY TASTE!
Yah! I'm so excited! :D
Flavors: Caramel, Fruity, Honey, Malt
i learned the bad news today. Steven Smith lost his battle with cancer. I love his teas.
This one is one of my favorite Yunnan Black( Red ) tea.
i brewed it Western today. my usual parameters 3g 8oz 200F for 3min
This tea is wonderful. its smooth and sweet but not overly. Im getting tired of sweet potato notes. thankfully this is not this kind. Some floral and savory. very enjoyable cup.


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10 of 11 people found the following review helpful
This review is from: Brahms: Ein Deutsches Requiem (German Requiem) (SDG: SDG706) (Audio CD)
John Eliot Gardiner seems to polarise people - folk seem either to love him or hate him. I know him from his Bach cantata recordings, having collected the entire set. But Brahms? I enjoyed his symphonies by Gardiner, but I know little about the Deutsches Requiem, so this was a speculative purchase. I enjoyed it thoroughly. A recording of a live performance, as are most Gardiner recordings, I found it quite thrilling. The Schütz choral pieces that are also on the CD are also excellent. The Monteverdis and the Big Gardiner Band are in top form. I can't compare it with anyone else's Deutsches Requiem, because I haven't heard anyone else's, but taken on its own, I found it really excellent both musically and performance-wise and would recommend a listen.
Review Details
(10 customer reviews)
Location: Switzerland
Top Reviewer Ranking: 1,033


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Arizona Cardinals
Looks like Arizona Cardinals' Antonio Cromartie is still fast
In 2006, as he was preparing to enter the NFL, Antonio Cromartie impressed the scouting world when he ran the 40-yard dash in 4.47 seconds.
At his size, that kind of speed was very, very impressive, and it helped lead him being selected 19th overall by the San Diego Chargers.
Cromartie has since put together a pretty impressive NFL career, picking off 28 passes over the course of eight seasons split between the Chargers and New York Jets.
He had a bit of a down year in 2013, though, intercepting just three passes and struggling at times in coverage. That could be attributed to a hip injury he was dealing with -- at least, that's what the Arizona Cardinals are banking on after signing the veteran to a one-year contract in the offseason.
Now 30, Cromartie is trying to prove he's still got it, and an Instagram video he posted Friday afternoon shows he's still plenty fast.
And ppl say I've lost a step. Only thing I can do laugh at you all. Ran 4.34 & 4.36 in the 40 today after a workout and I'm only getting faster thanks to @proactivesp and @goldmedalexcellence_chrisasher
Most Popular
Cardinals Audio
- Kurt Warner- NFL Network analyst - Tuesday March 31Kurt Warner describes what he saw at Jameis Winston's pro day.
- Daniel Jeremiah- NFL Network analyst - Friday March 27Daniel Jeremiah could see the Cardinals drafting a running back in a round after the first round.
- Kevin Guy, AZ Rattlers head coach - Friday March 27Coach Guy talks about losing players to the NFL and the challenges the Rattlers face this season.
- Charles Robinson, Yahoo! Sports NFL reporter - Tuesday March 24Charles Robinson believes that Adrian Peterson's relationship with the Vikings will only get worse.
- Sal Paolantonio- ESPN NFL analyst - Tuesday March 24Bruce Arians told Sal Paolantonio that there are "no serious discussions" about trading for Adrian


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Kingdom of the Sun
Set in ancient Egypt, this is a love story, tender and at the same time erotic and sensitive, that will enchant and captivate the readerwith its splendour of historical detail and explicit, unflinchingly sanguine portrayal of a time notorious for its savagery and cruelty. Even though she was a princess, Zarah was still a prisoner and had to act out the part that had been set for her. As Egypt's struggle for power swept on, a young girl, whose beauty was beyond price, had become a pawn in the game of treachery and intrigue played by Egypt's greatest queen. Zarah was plotting her revenge on those who had killed the only people she had ever cared for, but she was being watched by Per Ibsen. Was he just waiting his time, learning the truthuntil the moment he had her in his power?
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3 bedroom flats for sale in OX16
3 bed flat for saleMiddleton Road, Banbury OX16
Located on the popular Eastern side of Banbury is this highly unique and spacious three bedroom duplex apartment. Part of an attractive converted Victorian building, and with it's very own private garden, this charming property offers plenty of ... More details
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Marketed by Maxwells Estate Agents Ltd, OX16. Call 01295 675031
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Zoopla is one of the UK's leading property portals, helping you to find property for sale and to rent and make smarter decisions when buying and renting homes in the UK. Discover information on homes in OX16 by researching OX16 property values, OX16 house prices paid, our OX16 property market overview and find OX16 agents.


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Launcher Update
Get the Desktop App for Battle.net Now
- All your games in 1 place
- Log in once
- Automatic game updates
Destroy 30 units with Infested Terrans in an Unranked or Ranked game.
Win 50 1v1 Unranked or Ranked games as Zerg.
Destroy 10 cloaked or burrowed units in a single Melee game.
Attack for 20 seconds with no counterattacks in an Unranked or Ranked game.
Have 21 Zerg units morphing simultaneously in an Unranked or Ranked game.


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Discussion in 'Stillwater' started by yellowlab, Apr 5, 2011.
Premature release-ulation, It haunts me.
Nice one boot!
"Wait!! Come back!! It is a "catch and release" net!! Eeeeek!"
"Already got a tripod, now if I can only land this trploid."
Congratulations to Matt Baerwalde, and thanks for playing along! Hope we see some more photos and threads like these in the future.


text: Share: Diabetic Desserts Looking for diabetic desserts? Find delicious diabetic desserts including cookies, cakes, and more desserts for diabetics. Recipe Spotlight Pumpkin Oatmeal Bars It took me a long time to develop this quick-and-easy recipe, but I'm so happy with how it turned out. These bars have it all—sugar and spice and a light, creamy-rich pumpkin layer that's especially nice! —Erin Andrews, Edgewater, Florida View Pumpkin Oatmeal Bars Recipe Sign up for Our Recipeof the Day Newsletter New recipes and fan favorites delivering daily inspiration! SIGN UP More Diabetic Desserts Sort: Highest Rated Alphabetical Newest < prev | ... | 6 | 7 | 8 | 9 | 10 | ... | next > Icebox Sandwiches Icebox Sandwiches My mother liked making these cool creamy treats when I was growing up in the States because they're so quick to fix. Now my three kids enjoy them. View Recipe Little Orange Dream Cups Little Orange Dream Cups Our Test Kitchen used a combination of fat-free and reduced-fat dairy products in these creamy orange-flavored cups. Try substituting gingersnaps for the chocolate wafers to create a super-crisp variation. View Recipe No-Guilt Brownies No-Guilt Brownies Even those watching their diet can indulge in these light but luscious brownies.—Rita Ross, Delta, Ohio View Recipe Advertise with us ADVERTISEMENT Pineapple Delight Pineapple Delight I received this recipe at a meeting for diabetics many years ago. I've shared this dessert with many friends on restricted diets. View Recipe Quick Peanut Butter Pudding Quick Peanut Butter Pudding Joyce Crouse of Chambersburg, Pennsylvania dresses up sugar-free instant pudding with peanut butter for this simply yummy dessert. "I came up with the ides one day when I was looking for something fast for dessert. It's delicious," assures Joyce. View Recipe Strawberry Dessert Strawberry Dessert This recipe yields a large volume, so it's great to take to a potluck or family gathering. The cool, creamy pudding pairs well with the angel food cake and strawberries. View Recipe Tart 'n' Frosty Limeade Dessert Tart 'n' Frosty Limeade Dessert Luscious with lime juice, this frozen treat is so lovely and light that it melts all resistance to dessert—even after hearty holiday meals! Thanks to Pewaukee, Wisconsin's Pam Peirce for sharing a real winner. View Recipe Fluffy Key Lime Pie Fluffy Key Lime Pie For a taste of paradise, try this light and creamy confection. It's low in fat, sugar and fuss. Dessert doesn't get any easier! —Frances VanFossan, Warren, Michigan View Recipe Pineapple Pudding Cake Pineapple Pudding Cake My mother used to love making this easy dessert in the summertime. It's so cool and refreshing that it never lasts very long! Kathleen Worden - North Andover, Massachusetts View Recipe Yummy Chocolate Cake Yummy Chocolate Cake My husband and I are trying to eat lighter but still crave sweets. This moist, chocolate cake really helps with that. With the rich frosting, it makes a decadent treat! —LaDonna Reed, Ponca City, Oklahoma View Recipe White Chocolate Cranberry Cookies White Chocolate Cranberry Cookies These sweet cookies feature white chocolate and cranberries for a delightful taste. And the red and white coloring add a great holiday feel to any cookie tray. —Donna Beck, Scottdale, Pennsylvania View Recipe < prev | ... | 6 | 7 | 8 | 9 | 10 | ... | next > Advertise with us ADVERTISEMENT Advertise with us ADVERTISEMENT


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