An adaptable multiple power source for mass spectrometry and other scientific instruments.

An Adaptable Multiple Power Source (AMPS) system has been designed and constructed. The AMPS system can provide up to 16 direct current (DC) (±400 V; 5 mA), 4 radio frequency (RF) (two 500 VPP sinusoidal signals each, 0.5-5 MHz) channels, 2 high voltage sources (±6 kV), and one ∼40 W, 250 °C temperature-regulated heater. The system is controlled by a microcontroller, capable of communicating with its front panel or a computer. It can assign not only pre-saved fixed DC and RF signals but also profiled DC voltages. The AMPS system is capable of driving many mass spectrometry components and ancillary devices and can be adapted to other instrumentation/engineering projects.

Read-noise characterization of focal plane array detectors via mean-variance analysis.

Mean-variance analysis is described as a method for characterization of the read-noise and gain of focal plane array (FPA) detectors, including charge-coupled devices (CCDs), charge-injection devices (CIDs), and complementary metal-oxide-semiconductor (CMOS) multiplexers (infrared arrays). Practical FPA detector characterization is outlined. The nondestructive readout capability available in some CIDs and FPA devices is discussed as a means for signal-to-noise ratio improvement. Derivations of the equations are fully presented to unify understanding of this method by the spectroscopic community.

Effect of x-irradiation on growth and the expression of parathyroid hormone-related peptide and Indian hedgehog in the tibial growth plate of the rat.

AIM: To study the effect of irradiation on the longitudinal growth and the expression of parathyroid hormone-related peptide (PTHrP) and Indian hedgehog (IHh) in tibial growth plates of rats. METHODS: At 3 weeks of age, 30 male rats received a single fraction of irradiation (8 Gy) to their right hind limb, and small groups of animals were sacrificed 1, 2, 3, 5, 7, 10, 15, and 26 weeks after irradiation. Weight and length of both irradiated and nonirradiated tibiae were measured, and sections of the tibiae were stained with HE. PTHrP and IHh were visualized using immunohistochemical techniques. RESULTS: Radiation resulted in persistent growth delay of the irradiated tibiae, with a difference in length of more than 10% between the irradiated and the nonirradiated tibiae 15 weeks or more after irradiation. The growth plate architecture was disturbed, and the expression of both PTHrP and IHh was decreased in the irradiated tibiae. CONCLUSION: As PTHrP and IHh are key regulators of both the pace and the synchronization of the differentiation of growth plate chondrocytes, the reduced expression of PTHrP and IHh may contribute to the changes found after irradiation.

A sensitive assay for studying dopaminergic activity in cultures of rat pituitary cells.

The dopaminergic and antidopaminergic activity of drugs is frequently assayed in pituitary cell cultures. Here we describe a modified version of the assay based on the use of pituitary cells from prepubertal female rats. Under our experimental conditions (50,000 cells well-1, 2-day culture and 2-h drug-exposure) the assay yielded high selectivity and sensitivity for drug dopaminergic activity. D2 agonistic activity of bromocriptine could be observed at a concentration as low as 10(-15) M, the antagonistic activity of haloperidol at 10(-16) M. The assay also proved reproducible and simple enough for routine screening of dopaminergic activity. The assay enabled dopaminergic agonist and antagonist activity to be revealed at very low drug concentrations. The high sensitivity of the assay could be of advantage in studying dopaminergic activity in samples containing active substances present at low concentrations or for disclosing the activity of substances with low dopaminergic potency.

Analytical atomic spectroscopy using ion trap devices.

Investigations using ion trap devices for analytical atomic spectroscopy purposes have focused on the use of an inductively coupled plasma (ICP) ion source with ion trap mass spectrometric (ITMS) detection. Initial studies were conducted with an instrument assembled by simply appending an ion trap as the detector to a fairly conventional ICP/MS instrument, i.e. leaving an intermediate linear quadrupole between the plasma source and the ion trap. The principal advantages found with this system include the destruction of nearly all problematic and typical ICP/MS polyatomic ions (e.g., ArH(+), ArO(+), ArCl(+), Ar(2)(+), etc) and a dramatic reduction of the primary plasma source ion, Ar(+). These results prompted the development of a second-generation plasma source ion trap instrument in which direct coupling of the ICP and ion trap has been effected (i.e. no intermediate linear quadrupole); the same performance benefits have been largely preserved. Initial operation of this instrument is described, characterized, and compared to the originally described ICP/ITMS and conventional ICP/MS systems. In addition, experiments aimed at improving ICP/ITMS sensitivity and selectivity using broadband resonance excitation techniques are described. Finally, the potential for laser optical detection of trapped ions for analytical purposes is speculated upon.

Plasma source ion trap mass spectrometry: Enhanced abundance sensitivity by resonant ejection of atomic ions.

An experimental study of resonant ion excitation in an rf quadrupole ion trap is reported. Atomic ions are generated in an inductively coupled plasma and injected into the ion trap where, after collisional cooling, they are irradiated by a low-voltage, dipole coupled waveform. Single frequency, narrowband, and broadband excitation pulses have been used. Absorption lineshapes (plots of observed ion signal versus excitation frequency) are shown for variations in buffer gas pressure and the amplitude and duration of the single frequency pulses. The absorption lineshapes are usually asymmetric and tail toward lower frequencies. At sufficiently low buffer gas pressure or potential well depth, the lineshapes broaden and become more asymmetric to the point that absorption by ions with adjacent mass-to-charge ratios overlaps. This overlapping absorption reduces the selectivity with which a single mass-to-charge ratio ion can be excited and ejected relative to nearby mass-to-charge ratio ions. The rate of ion ejection is different on the low versus high frequency edges of the absorption lines. This difference in ejection rates provides an important key to understanding the shape of the absorption lines. All of these observations are explained in terms of the known kinematic behavior of ions in real traps, that is, traps with substantial higher order symmetry components in the trapping field ("nonlinear" fields). The importance of the nonlinearity of the trapping field in understanding the observed lineshapes and their time dependencies is discussed. We also report resonant ejection results obtained using multiple frequency (narrow or broad bandwidth) excitation. Multiple frequency excitation allows ions with different mass-to-charge ratio values to be ejected from the trap using one excitation waveform. The finite ion storage capacity of the ion trap is thereby reserved for the ion(s) of interest. We show that ejection of (89)Y ions can be ∼ 10(5) times more efficient than ejection of ions at either m/z 88 or 90.

On the dynamics between gonadotrophin surge-inhibiting factor and gonadotrophin releasing hormone (GnRH): role of self-priming and desensitization in the luteinizing hormone response to GnRH after follicle stimulating hormone treatment.

The effects were studied of follicle stimulating hormone (FSH)-induced production of gonadotrophin surge-inhibiting factor (GnSIF) on three phases of the pituitary responsiveness to gonadotrophin releasing hormone (GnRH): the unprimed, primed and desensitized phases. Rats were injected with FSH on two occasions during the oestrous cycle. Spontaneous luteinizing hormone (LH) surges were measured as well as GnRH-induced LH surges on the day of pro-oestrus during infusions with 100-4000 pmol GnRH/rat/10 h, in phenobarbital blocked rats. The spontaneous LH surges were attenuated or completely inhibited by the FSH treatment. FSH suppresses and prolongs the unprimed LH response and delays GnRH self-priming, especially during infusions with low concentrations of GnRH. This treatment does not affect the total LH response (area under curve) to the highest concentrations of GnRH and after ovariectomy. On the other hand, this response is suppressed during infusions with the lower concentrations of GnRH. Hence, FSH, via GnSIF, delays maximal priming of the LH response to GnRH, whereas the suppression of LH release is a consequence of the GnRH-induced progressed state of desensitization. The inconsistent effects of FSH on the mid-cycle LH surges are explained as a result of the interaction between the relative strengths of GnRH and GnSIF.

Induction of the gonadotrophin surge-inhibiting factor by FSH and its elimination: a sex difference in the efficacy of the priming effect of gonadotrophin-releasing hormone on the rat pituitary gland.

This study was designed to explore the efficacy of gonadotrophin-releasing hormone (GnRH) to antagonize the effect of gonadotrophin surge-inhibiting factor (GnSIF) on the timing of the induction by GnRH of the maximal self-priming effect on pituitary LH responsiveness. The GnSIF levels were increased by FSH treatment and reduced after gonadectomy. Female rats were injected s.c. with 10 IU FSH or saline (control) on three occasions during the 4-day cycle. Serial i.v. injections of GnRH (500 pmol/kg body weight) were administered to intact rats on the afternoon of pro-oestrus or 15-30 min after ovariectomy. Intact male rats were given 10 IU FSH and 500 or 2000 pmol GnRH/kg body weight on an equivalent time-schedule. Endogenous GnRH release was suppressed with phenobarbital. In intact female control rats, the timing of the maximally primed LH response was delayed as the GnRH pulse-interval increased. FSH treatment of female rats induced a suppression of the initial unprimed LH response and delayed the maximally primed LH response, which showed further delay as the GnRH pulse-interval was increased. When the pulsatile administration of GnRH was started 15-30 min after ovariectomy, the priming effect of GnRH did not change as the GnRH pulse-interval was increased in the saline-treated rats. However, FSH treatment caused a suppression of the unprimed LH response, a delay in the primed LH response and decreased the delay of the maximally primed LH response to GnRH when the GnRH pulse-interval was decreased. Increasing the interval between ovariectomy and the first GnRH pulse to 4 h diminished the efficacy of the FSH treatment: GnRH-induced priming was delayed by only one pulse instead of the two pulses in control rats. In intact males but not in orchidectomized rats, a self-priming effect was demonstrated during GnRH pulses which were 1 h apart. The effect of 2 nmol GnRH/kg body weight was the most pronounced. Compared with intact female rats, the timing of the maximally primed LH response was delayed by 1 h. FSH treatment did not affect the pituitary LH response to both dose levels of GnRH. It is concluded that FSH treatment increased the release of GnSIF by the ovary, then induced a state of low responsiveness of the pituitary gland to GnRH and subsequently delayed GnRH-induced maximal self-priming. The efficacy of GnRH to prime the pituitary gland was higher when GnSIF levels were decreasing after removal of the ovaries.(ABSTRACT TRUNCATED AT 400 WORDS)

The effect of gonadotrophin surge-inhibiting factor on the self-priming action of gonadotrophin-releasing hormone in female rats in vitro.

The present study was designed to explore further the functional antagonism between gonadotrophin-releasing hormone (GnRH) and the ovarian factor, gonadotrophin surge-inhibiting factor (GnSIF). In all experiments, pituitary tissue was exposed to various amounts of GnSIF, after which the self-priming action of GnRH was studied. GnSIF was increased in vivo by FSH treatment and increased in vitro by adding various amounts of follicular fluid (FF) to cultured pituitary cells. Treatment with 3 or 10 IU FSH suppressed the initial LH response and delayed the maximally primed LH response to GnRH. Treatment with FSH was only effective in intact rats on days 1 and 2 of dioestrus. There was no difference in the rate of maximal LH release irrespective of treatment with either FSH or saline. Since FSH treatment was ineffective in long-term ovariectomized rats, it was concluded that the initial suppressive effect of FSH on LH release was mediated by GnSIF. Cycloheximide prevented the self-priming action of GnRH by inhibiting GnRH-induced protein synthesis. The initial protein synthesis-independent GnRH-stimulated LH release, which was already suppressed by FSH treatment, remained suppressed in the presence of cycloheximide. Pretreatment with GnRH in vivo increased the protein synthesis-independent GnRH-induced LH release during subsequent incubation of the glands. This increase did not occur after FSH treatment. Pituitary cells, cultured for 20 h in medium only, failed to elicit the self-priming effect of GnRH. Preincubation with FF maintained the self-priming effect. This was independent of the concomitant presence of various amounts of oestradiol. Preincubation with bovine FF suppressed the initial GnRH-stimulated LH release dose-dependently. Porcine FF, human FF and testicular extract suppressed the release of LH in a similar way. It was concluded that GnSIF suppresses the initial LH response to continuous GnRH stimulation. Increased levels of GnSIF caused by FSH treatment also delayed the primed LH release. The mechanism of functional antagonism between GnSIF and GnRH could give rise to the occurrence of the phenomenon of GnRH self-priming.

The self-priming action of LHRH is under negative FSH control through a factor released by the ovary: observations in female rats in vivo.

Female rats were treated with Metrodin (highly purified urinary FSH from menopausal women) or saline during the oestrous cycle. On the day of pro-oestrus they were anaesthesized with phenobarbital and received four repetitive LHRH injections 1 h apart. This treatment with FSH suppressed the unprimed LH response to the first LHRH injection. During the subsequent injections the maximal LHRH self-priming was delayed by 3 h till the fourth LHRH stimulation. At this time, LH release in response to LHRH was equally as high as shown in the saline controls after the second LHRH injection. Ovariectomized rats did not show the self-priming effect and FSH treatment was ineffective in suppressing LHRH-induced LH release. Administration of FSH followed by an additional 4- or 24-h period before LHRH stimulation were equally effective in suppressing the unprimed LH release and delaying (up to 3 h) the maximal priming of LH release by LHRH. Even 4-20-fold increased amounts of LHRH did not affect the suppressed unprimed release of LH after FSH treatment. Treatment with FSH did not change oestradiol and progesterone levels. It was concluded that FSH treatment suppresses the unprimed LHRH-induced LH release and delays maximal LHRH self-priming by enhancing the release of an ovarian factor.

Effects of AF64A on [3H]acetylcholine synthesis in neuron-enriched primary brain cell cultures.

[3H]Acetylcholine (ACh) synthesis was measured in primary neuronal cultures from neonatal rat brains. Neuronal [3H]ACh synthesis was blocked by hemicholinium-3 and depended on the age of the cultures, increasing for ca. 10 days, and eventually declining. The irreversible inhibitor AF64A (10 or 30 microM) inhibited [3H]ACh synthesis from [3H]choline at concentrations (10 or 30 microM) with affecting choline acetyltransferase activity. Nine-day-old cultures recovered 90% of their [3H]ACh synthesis within 7 days after AF64A, while 13-day-old cultures never recovered. These results suggest that the turnover of neuronal choline transporters is age-related.