The rapid immunosuppression in phytohemagglutinin-activated human T cells is inhibited by the proliferative Ca(2+) influx induced by progesterone and analogs.

Progesterone, an endogenous immunomodulator, suppresses human T-cell activation during pregnancy. A sustained Ca(2 +) influx is an important signal for T-cell proliferation after crosslinking of T-cell receptor/CD3 complexes by anti-CD3 antibodies or phytohemagglutinin (PHA). Progesterone targets cell membrane sites inducing rapid responses including elevated intracellular free calcium concentration ([Ca(2+)]i) and suppressed T-cell PHA-activated proliferation. Interestingly, both PHA and progesterone induce [Ca(2+)]i elevation, but it remains unclear whether the PHA-induced Ca(2+) influx is affected by progesterone leading to T-cell immunosuppression. Primary T-cells were isolated from human peripheral blood and the quench effect on intracellular fura-2 fluorescence of Mn(2+) was used to explore the responses to Ca(2+) influx with cell proliferation being determined by MTT assay. PHA-stimulated Ca(2+) influx was dose-dependently suppressed by progesterone and its agonist R5020, which correlated with PHA-activated T-cell proliferation inhibition. A similar dose-dependent suppression effect on cellular Ca(2+) influx and proliferation occurred with the TRPC channel inhibitor BTP2 and selective TRPC3 channel inhibitor Pyr3. In addition, two progesterone analogs, Org OD 02-0 and 20α-hydroxyprogesterone (20α-OHP), also produced dose-dependent suppression of Ca(2+) influx, but had no effect on proliferation. Finally, inhibition of PHA-activated T-cell proliferation by progesterone is further suppressed by 20α-OHP, but not by Org OD 02-0. Overall, progesterone and R5020 are able to rapidly decrease PHA-stimulated sustained Ca(2+) influx, probably via blockade of TRPC3 channels, which suppresses T-cell proliferation. Taken together, the roles of progesterone and its analogs regarding the rapid response Ca(2+) influx need to be further explored in relation to cytokine secretion and proliferation in activated T-cells.

Non-genomic rapid responses via progesterone in human peripheral T cells are not indirectly mimicked by sphingosine 1-phosphate.

Progesterone is an endogenous immunomodulator that suppresses T cell activation during pregnancy. Progesterone has been shown to induce rapid responses that cause intracellular calcium ([Ca(2+)]i) elevation and acidification followed by inhibition of phytohemagglutinin (PHA)-stimulated proliferation. These rapid responses involve T cell plasma membrane sites, but the mechanisms remain unclear. Three new membrane progesterone receptors (mPRα/mPRß/mPRγ) have been identified as expressed in T cells. These proteins have been identified as G-protein-coupled receptors. Recently, mPRs have been classified as progestin and adipoQ receptors (PAQRs). Furthermore, they have been suggested to be alkaline ceramidases, possibly involved in mediating sphingolipid signaling. Alkaline ceramidases are capable of converting ceramide to sphingosine, which might then be further phosphorylated sphingosine via sphingosine kinase to sphingosine 1-phosphate (S1P). This pathway could result in progesterone acting indirectly via S1P on membrane sphingosine 1-phosphate receptors (S1PRs) in T cells to induce rapid responses. Therefore, our aim was to investigate whether progesterone rapid responses occur indirectly in T cells via S1P. We found that S1P induces [Ca(2+)]i elevation however there was no change in intracellular pH. This is different from the situation with progesterone: S1P alone does not suppress PHA-stimulated cell proliferation and does not act synergistically with progesterone on the inhibition of PHA-induced cell proliferation. In contrast, S1P at 1µM is able to antagonize the proliferation inhibitory effect of progesterone. Thus the rapid responses that are induced by progesterone in human peripheral T cells probably do not involve indirect signaling via S1P and S1PRs.

A new on-chip all-digital three-phase full-bridge dc/ac power inverter with feedforward and frequency control techniques.

The communication speed between components is far from satisfactory. To achieve high speed, simple control system configuration, and low cost, a new on-chip all-digital three-phase dc/ac power inverter using feedforward and frequency control techniques is proposed. The controller of the proposed power inverter, called the shift register, consists of six-stage D-latch flip-flops with a goal of achieving low-power consumption and area efficiency. Variable frequency is achieved by controlling the clocks of the shift register. One advantage regarding the data signal (D) and the common clock (CK) is that, regardless of the phase difference between the two, all of the D-latch flip-flops are capable of delaying data by one CK period. To ensure stability, the frequency of CK must be six times higher than that of D. The operation frequency of the proposed power inverter ranges from 10 Hz to 2 MHz, and the maximum output loading current is 0.8 A. The prototype of the proposed circuit has been fabricated with TSMC 0.35 µm 2P4M CMOS processes. The total chip area is 2.333 x 1.698 mm2. The three-phase dc/ac power inverter is applicable in uninterrupted power supplies, cold cathode fluorescent lamps, and motors, because of its ability to convert the dc supply voltage into the three-phase ac power sources.

Low-sensitivity, low-bounce, high-linearity current-controlled oscillator suitable for single-supply mixed-mode instrumentation system.

A low-sensitivity, low-bounce, high-linearity current-controlled oscillator (CCO) suitable for a single-supply mixed-mode instrumentation system is designed and proposed in this paper. The designed CCO can be operated at low voltage (2 V). The power bounce and ground bounce generated by this CCO is less than 7 mVpp when the power-line parasitic inductance is increased to 100 nH to demonstrate the effect of power bounce and ground bounce. The power supply noise caused by the proposed CCO is less than 0.35% in reference to the 2 V supply voltage. The average conversion ratio KCCO is equal to 123.5 GHz/A. The linearity of conversion ratio is high and its tolerance is within +/-1.2%. The sensitivity of the proposed CCO is nearly independent of the power supply voltage, which is less than a conventional current-starved oscillator. The performance of the proposed CCO has been compared with the current-starved oscillator. It is shown that the proposed CCO is suitable for single-supply mixed-mode instrumentation systems.

An active current-sensing constant-frequency HCC buck converter using phase-frequency-locked techniques.

A hysteresis-current-controlled (HCC) buck converter with active current-sensing and phase-frequencylocked techniques is presented in this paper. The proposed active current-sensing technique can not only consume less power than previous techniques, but also fully sense the inductor current. Although the buck converter is HCC, the switching frequency can be constant due to the devised phase-frequency-locked technique. The proposed converter has been designed and implemented with TSMC 0.35 microm DPQM CMOS processes. It is shown in the experimental results that the HCC buck converter features the following characteristics: 1) up to 800 mA of load current, 2) wide input and output voltage range, 3) high power efficiency, and 4) constant-frequency operation.

Direct effect of propylthiouracil on progesterone release in rat granulosa cells.

1. The present study was to investigate the direct effect and action mechanism of propylthiouracil (PTU), an antithyroid drug, on the production of progesterone in rat granulosa cells. 2. PTU (3-12 mM) decreased the basal and human chorionic gonadotropin (hCG)-stimulated release of progesterone from rat granulosa cells. 3. PTU (3-12 mM) attenuated the stimulatory effects of forskolin and 8-bromo-cyclic 3':5'-adenosine monophosphate on progesterone release from rat granulosa cells. 4. PTU (12 mM) inhibited the activities of both the cytochrome P450 side-chain cleavage enzyme (P450scc, conversion of 25-hydroxyl cholesterol to pregnenolone) and the 3beta-hydroxysteroid dehydrogenase (conversion of pregnenolone to progesterone) in rat granulosa cells. PTU decreased the V(max) but increased the K(m) of P450scc. 5. PTU (12 mM) decreased the hCG-increased amount of steroidogenic acute regulatory (StAR) protein in rat granulosa cells. 6. The present results suggest that PTU decreases the progesterone release by granulosa cells via a thyroid-independent mechanism involving the inhibition of post-cAMP pathway, and the activities of intracellular calcium, steroidogenic enzyme, and StAR protein functions.

Progesterone attenuates the inhibitory effects of cardiotonic digitalis on pregnenolone production in rat luteal cells.

Previous studies have shown that digoxin decreases testosterone secretion in testicular interstitial cells. However, the effect of digoxin on progesterone secretion in luteal cells is unclear. Progesterone is known as an endogenous digoxin-like hormone (EDLH). This study investigates how digitalis affected progesterone production and whether progesterone antagonized the effects of digitalis. Digoxin or digitoxin, but not ouabain, decreased the basal and human chorionic gonadotropin (hCG)-stimulated progesterone secretion as well as the activity of cytochrome P450 side chain cleavage enzyme (P450scc) in luteal cells. 8-Br-cAMP and forskolin did not affect the reduction. Neither the amount of P450scc, the amount of steroidogenic acute regulatory (StAR) protein, nor the activity of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) was affected by digoxin or digitoxin. Moreover, in testicular interstitial and luteal cells, progesterone partially attenuated the reduction of pregnenolone by digoxin or digitoxin and the progesterone antagonist, RU486, blocked this attenuation. These new findings indicated that (1) digoxin or digitoxin inhibited pregnenolone production by decreasing the activity of P450scc enzyme, but not Na(+)-K(+)-ATPase, resulting in a decrease on progesterone secretion in rat luteal cells, and (2) the inhibitory effect on pregnenolone production by digoxin or digitoxin was reversed partially by progesterone. In conclusion, digoxin or digitoxin decreased progesterone production via the inhibition of pregnenolone by decreasing P450scc activity. Progesterone, an EDLH, could antagonize the effects of digoxin or digitoxin in luteal cells.