Cold defect on indium leukocyte scanning of a hepatic abscess.

Abdominal and, specifically, hepatic abscesses have always been difficult to diagnose clinically. The In-111 leukocyte scan has proven to be a valuable modality assisting in the diagnosis of an abscess. The authors present the case of a 64-year-old man who presented with vague clinical symptoms and inconclusive laboratory data for a hepatic abscess. Both the ultrasound and CT scan were nondiagnostic; however, a distinct cold defect within the liver on an In-111 leukocyte scan resulted in a liver biopsy, confirming the presence of an abscess. The potential causes of cold defects on In-111 WBC scanning and the limitations of this modality are discussed.

Endocrine activity of induced persistent follicles in sheep.

This experiment was designed to examine gonadotropin requirements for the induction and maintenance of persistent ovarian follicles in sheep. At the time of prostaglandin (PG) treatment on the tenth day of an induced estrous cycle, 8 ewes (with one ovary autotransplanted to the neck) received an injection of a GnRH antagonist ([Ac-d-Nal1, d-4-C-1-Phe2, d-Trp3, d-Arg6, d-Ala10] GnRH.HOAc; 50 microg/kg s.c.), and continuous hourly injections of exogenous ovine LH (equivalent to 1.25 microg NIH-oLH-S26) began simultaneously with this first antagonist injection (time zero). Antagonist was given three times at 3-day intervals. On Day 6, LH injections were stopped in 4 ewes (group 2) but continued in 4 other ewes (group 1) until the end of the 10-day experiment. Ovarian vein blood was sampled daily every 15 min for a 2-h period around two injections of exogenous LH (this sampling included group 2 after Day 6). Additional jugular and ovarian vein blood samples were collected every 8 h throughout the experiment. Daily ultrasound examination revealed the presence of at least one large follicle (range 4- to 7.5-mm diameter) from Day 3 to Day 10 in all ewes, but no new growing follicles (> 2 mm) were detected for at least 6 days. After Day 2, secretion of estradiol was positively correlated with that of inhibin (r = 0.83, p < 0.001), whereas FSH concentrations were inversely related to inhibin (r = -0.71, p < 0.001) and estradiol (r = -0.81, p < 0.001). In the absence of an LH surge, estradiol and androstenedione secretion (range 5-20 ng steroid/min) was maintained from Day 1 to Day 8 in group 1; but in group 2, secretion decreased abruptly when the LH injections stopped. Thus, continued low-amplitude, high-frequency LH pulses were required to maintain estradiol secretion when concentrations of FSH were < 0.5 ng/ml. However, estradiol and androstenedione secretion decreased (and FSH concentrations increased) between Days 8 and 10 in the ewes that received continued LH injections (group 1), showing that atresia in estrogenic follicles was not due to a lack of gonadotropin availability but to changes within the follicle. For the first 3 days after administration of PG, androstenedione secretion was greater than that of estradiol (p < 0.05), but from Day 4 to 6 the secretion rates were similar (p < 0.1), suggesting that aromatase may be limiting in the first 3 days whereas provision of androstenedione precursors was altered as the follicle persisted. In group 2 on Days 7 and 8 when hourly LH injections had stopped, neither androstenedione nor estradiol secretion increased after one test injection of LH; in contrast, androstenedione but not estradiol secretion increased after a second LH test injection 1 h later, suggesting that secretion of androstenedione is controlled by repeated exposure to LH. In conclusion, persistent estrogenic follicles were produced in the follicular phase in sheep by treatment with a combination of GnRH antagonist and hourly pulses of LH. Secretion of estradiol was dependent on continued hourly LH pulses of approximately 1 ng/ml and the follicles remained estrogenic for 8 days, after which time the ability to secrete estradiol and androstenedione declined even with continued LH injections.

Recurrent lower gastrointestinal hemorrhage: ileal neoplasm diagnosed by scintigraphy with Tc 99m red blood cells and angiography.

A 71-year-old man with recurrent lower gastrointestinal bleeding had negative colonoscopic findings on three occasions. Preoperative diagnosis of a large stromal tumor in the right lower quadrant was based on scintigraphy with tagged red blood cells and mesenteric arteriographic features. The use of nuclear scintigraphy in the diagnosis and evaluation of stromal tumor of unknown malignant potential has not been previously described.

Ectopic adrenal myelolipoma presenting as a splenic mass.

Myelolipoma is a benign tumor composed of mature adipose tissue and hematopoietic elements. Although they are most commonly found in the adrenal glands, extra-adrenal myelolipomas are well documented. We describe a case of myelolipoma arising within or adjacent to the spleen of a patient with sickle cell disease. This encapsulated tumor contained myeloid and adipose components, as well as clear cells resembling adrenal cortical tissue. Although ectopic adrenal tissue has not been reported in the spleen, we feel that this is the most likely site of origin of this mass. To our knowledge, this is the first instance of this neoplasm presenting as a splenic mass. The possible pathogenesis and etiology of this unusual lesion is discussed.

The effect of acute immuno-neutralisation of inhibin in ewes during the early luteal phase of the oestrous cycle on ovarian hormone secretion and follicular development.

Ewes with ovarian autotransplants received either inhibin antiserum (10 ml i.v. raised in sheep against recombinant 32 kDa human inhibin; n = 6) or sheep serum (10 ml i.v.; n = 5) on day 3 of the luteal phase with additional daily injections (1 ml i.v.) from 48 h after the initial bolus until day 13. Jugular and ovarian venous blood samples were taken 4-hourly over days 2-13 of the luteal phase. Blood samples were also taken at more frequent intervals (every 10-15 min for 2-3 h) to examine pulsatile secretory responses from the ovary to endogenous and gonadotrophin-releasing hormone-induced (150 ng i.m.) LH pulses on days 4, 6, 8, 10 and 12 of the luteal phase. Plasma FSH levels, ovarian steroid secretion and ovarian follicular development were measured. The ovarian follicle population was estimated daily by real time ultrasound scanning. Immunisation against inhibin resulted in a 3- to 4-fold increase (P < 0.001) in plasma FSH levels within 8 h with levels remaining elevated over controls for 6-7 days. Within 24 h of immunisation there was an increase in the number of small ovarian follicles (P < 0.05) and by 3 days after treatment immunised ewes had 4-6 large ovarian follicles/ewe with this increase in the total number of large follicles being maintained for the rest of the experimental period (P < 0.05). Mean ovarian oestradiol secretion during intensive bleeds was not different from controls 24 h after immunisation, but by 3 days after immunisation it was elevated 4- to 5-fold (P < 0.001) over controls with this increase being maintained throughout the experiment. Similar responses to immunisation against inhibin in androstenedione secretion were observed although mean androstenedione secretion was not elevated until 7 days after treatment. In vitro antibody titres in immunised ewes remained elevated but declined steadily (P < 0.001) over the experimental period. We conclude that the initial stimulation of follicle development and ovarian steroid secretion following passive immunisation against inhibin can be attributed to increased blood FSH. However, the fact that with time FSH declined but increased follicle development was sustained, despite maintenance of high circulating antibody titres, suggests that on a longer term basis inhibin immunisation may stimulate ovarian function by interfering with the modulation of follicle development by inhibin at an ovarian level.

An analysis of the safety of performing dobutamine stress echocardiography in an ambulatory setting.

Dobutamine echocardiography has become widely used in the past decade in the evaluation of patients with suspected coronary artery disease who are unable to undergo exercise treadmill or bicycle testing. The safety of this procedure has been studied in a hospital-based setting. However, no studies thus far have evaluated the safety of this procedure in an office-based setting, remote from a hospital. We performed dobutamine echocardiography on 127 patients in an office-based setting, remote from a hospital. Throughout the course of this study there were no deaths, myocardial infarctions, sustained episodes of ventricular tachycardia, or syncopal episodes associated with dobutamine infusion. The frequency of noncardiac side effects was 29%, the majority of which were nausea, vomiting, and paresthesias. Three patients had nonsustained ventricular tachycardia, none of whom had symptoms. We conclude that dobutamine echocardiography is safe, well tolerated, and useful in an office-based setting.

The effect of ovarian arterial infusion of transforming growth factor alpha on ovarian follicle populations and ovarian hormone secretion in ewes with an autotransplanted ovary.

Transforming growth factor alpha (TGF alpha) inhibits hormone production by cultured follicular cells but evidence of an effect of TGF alpha on ovarian hormone secretion in vivo is still required. Eleven ewes with an autotransplanted ovary received, by ovarian arterial infusion, either 5 micrograms/h recombinant rat TGF alpha (n = 6) or placebo (n = 5) for 12 h on day 10 of the luteal phase. Two hours before the start and 1 hour before the end of the infusion each ewe received a single injection of gonadotrophin-releasing hormone (GnRH; 150 ng i.v.). Two hours after the end of the infusion luteal regression was induced with prostaglandin F2 alpha (PGF2 alpha; 125 micrograms i.m.). Ovarian and jugular venous blood samples were taken at 10-min, 15-min or 4-h intervals from 2 h before the start of the infusion until 96 h after PGF2 alpha and the rates of secretion of ovarian oestradiol, inhibin, progesterone and androstenedione were determined. Jugular venous concentrations of LH and FSH were also measured and follicle populations monitored by real-time ultrasound scanning. Infusion of TGF alpha resulted in a significant (P < 0.05) depression in the amplitude of the pulsatile response of oestradiol and androstenedione secretion to the GnRH-induced LH pulse at the end of the infusion. Ovarian inhibin secretion was acutely suppressed by TGF alpha infusion (P < 0.001) and remained lower than controls for the period of the experiment. Luteal phase progesterone secretion was also acutely inhibited (P < 0.001) by infusion of TGF alpha, and in one treated ewe progesterone secretion was elevated 48-84 h after PGF2 alpha. Jugular venous concentrations of FSH in TGF alpha-treated ewes were significantly (P < 0.001) elevated over controls during the first 48 h of the follicular phase and the LH surge was delayed for about 10 h (P < 0.05). Infusion of TGF alpha caused a marked decline (P < 0.05) in the number of large follicles within 12 h of the end of the infusion. Two of the six treated ewes, including the one with high follicular phase progesterone, had unusually large (8.7 and 10 mm) follicles present from 48-96 h after PGF2 alpha. We conclude that direct arterial infusion of TGF alpha results in acute inhibition of ovarian steroid and inhibin secretion that is associated with induction of atresia in the population of large follicles. The lack of feedback of ovarian hormones results in a rebound increase of FSH which stimulates the growth of more ovarian follicles and the eventual re-establishment of ovarian hormone secretion and normal cyclicity.

Effects of benzodiazepine receptor ligands on simultaneous visual discriminations of variable difficulty.

Benzodiazepine receptor (BZR) ligands have been demonstrated to affect the performance in tasks measuring attentional abilities. In such tasks, subjects typically are required to discriminate visual and/or auditory stimuli. The possibility that the effects of BZR ligands on the performance in tasks measuring attention are primarily due to effects on discriminative processes has not been tested systematically. Rats were trained to discriminate between simultaneously presented pairs of visual stimuli flashing either at 5 Hz versus 4.17, 3.75, 2.5, 1.67 or 1.25 Hz (group 1; FAST), or at 1.25, 1.46, 1.67, 2.5 or 3.33 Hz versus 5 Hz (group 2; SLOW) for 4.8 s (20 trials per discrimination type; sequence of pairs was randomized). In both groups, response accuracy depended significantly on the discriminability of the stimuli, with near perfect accuracy in response to most different pairs of stimuli and near chance-level accuracy in response to least different pairs of stimuli. Administration of the BZR full agonist chlordiazepoxide (1.56, 6.25, 9.38 mg/kg; i.p.) potently increased the number of errors of omission which, following the higher doses, confounded the effects on absolute numbers of correct and incorrect responses. However, the available data do not suggest that the agonist affected the animals' abilities to discriminate between the stimuli. Similarly, administration of the BZR ligands ZK 93 426 and MDL 26,479 (which negatively modulate GABAergic transmission) produced no systematic effects. These data suggest that the effects of BZR ligands on the performance in tasks measuring attentional abilities are not primarily due to effects on the animals' ability to discriminate sensory stimuli.

Purification of the endogenous glucocorticoid receptor stabilizing factor.

A ubiquitous, low molecular weight, heat-stable component of cytosol stabilizes the glucocorticoid receptor in its untransformed state in association with hsp90. This heat-stable factor mimics molybdate in its effects on receptor function, and it has the heat stability, charge, and chelation properties of a metal oxyanion [Meshinchi, S., Grippo, J.F., Sanchez, E.R., Bresnick, E.H., & Pratt, W.B. (1988) J. Biol. Chem. 263, 16809-16817]. In this paper, we describe the further purification of the endogenous factor from rat liver cytosol by anion-exchange HPLC (Ion-110) after prepurification by molecular sieving, cation absorption, and charcoal absorption. Elution of the factor with an isocratic gradient of ammonium bicarbonate results in recovery of all of the bioactivity in a single peak which coelutes with inorganic phosphate and contains all of the endogenous molybdenum. The bioactivity can be separated from inorganic phosphate by chromatography of the partially purified endogenous factor on a metal-chelating column of Chelex-100. The chelating procedure results in complete loss of bioactivity with recovery of 98% of the inorganic phosphate in both the column drop-through and a subsequent 1 M NaCl wash. The factor preparation purified through the Ion-110 HPLC step inhibits temperature-mediated dissociation of the immunopurified glucocorticoid receptor-hsp90 complex, but it is considerably more effective at stabilizing the unpurified receptor-hsp90 complex in a Chelex-treated cytosol system that has been depleted of metal components. These observations support the proposal that an endogenous metal can stabilize the binding of hsp90 to the receptor but it is likely that other cytosolic components that are not present in the immunopurified complex must contribute to the stability of the soluble protein-protein complex in cytosol.

Quantitative determination of selected compounds in a Kentucky 1R4F reference cigarette smoke by multidimensional gas chromatography and selected ion monitoring-mass spectrometry.

Eight compounds from a Kentucky 1R4F reference cigarette smoke condensate have been determined by selected ion monitoring-mass spectrometry (SIM-MS) to confirm the validity of multidimensional gas chromatography (MDGC) as a quantitative tool in complex mixture analyses. Four electrostatically precipitated smoke condensate samples of 100 cigarettes each are dissolved individually in 25 mL of 2-propanol. The 2-propanol contains two methyl esters (C8 and C14) and seven deuterium-labeled compounds used as internal standards (IS). Analysis of the compounds of interest, pyridine; acetamide; acrylamide; phenol; o-, m-, and p-cresol; and quinoline, is accomplished by using two heartcuts. Heartcut times of the MDGC analysis are selected such that at least one IS is transferred with each group of compounds being analyzed. This study shows that the MDGC technique previously developed and described can be used for quantitative analyses. A comparison is made between the two types of internal standards. The results obtained for both types of internal standards agree within 20% of each other, on the average, with higher standard deviations for approximately 60% of the compounds where methyl esters are used as internal standards.

An X-ray microprobe facility using synchrotron radiation.

An X-ray microprobe for trace elemental analysis at micrometer spatial resolutions, using synchrotron radiation (SR), is under development. The facility consists of two beamlines, one including a 1:1 focusing mirror and the other an 8:1 ellipsoidal mirror. At present, "white light" is used for excitation of the characteristic X-ray fluorescence lines. Sensitivities in thin biological samples are in the range of 2-20 fg in 100 microns2 areas in 5 min irradiation times. Scanning techniques, as well as microtomography and chemical speciation, are discussed. Application to a specific biomedical study is included.

Distribution of trace levels of therapeutic gallium in bone as mapped by synchrotron x-ray microscopy.

Gallium nitrate, a drug that inhibits calcium release from bone, has been proven a safe and effective treatment for the accelerated bone resorption associated with cancer. Though bone is a target organ for gallium, the kinetics, sites, and effects of gallium accumulation in bone are not known. We have used synchrotron x-ray microscopy to map the distribution of trace levels of gallium in bone. After short-term in vivo administration of gallium nitrate to rats, trace (nanogram) amounts of gallium preferentially localized to the metabolically active regions in the metaphysis as well as the endosteal and periosteal surfaces of diaphyseal bone, regions where new bone formation and modeling were occurring. The amounts measured were well below the levels known to be cytotoxic. Iron and zinc, trace elements normally found in bone, were decreased in amount after in vivo administration of gallium. These studies represent a first step toward understanding the mechanism(s) of action of gallium in bone by suggesting the possible cellular, structural, and elemental "targets" of gallium.

Analysis of flue-cured tobacco essential oil by hyphenated analytical techniques.

The major components of an alkaloid-free, flue-cured, tobacco essential oil sample are isolated and identified. This is accomplished by utilizing modern hyphenated analytical methods. The instrumentation developed to accomplish this are an automated multidimensional gas chromatograph/mass spectrometer/flame ionization detector (MDGC/MS/FID) and a multidimensional gas chromatograph/matrix isolation/Fourier transform infrared spectrometer (MDGC/MI/FTIR). A total of 306 compounds is identified in the essential oil, of which 80 are found as tobacco constituents for the first time.

X-ray fluorescence with synchrotron radiation.

The use of synchrotron radiation for X-ray fluorescence has several advantages over the use of other conventional X-ray sources. The principles of synchrotron radiation and methods for applying synchrotron radiation to the X-ray fluorescence measurements of trace elements are discussed. The Brookhaven National Laboratory X-ray microprobe, facilities dedicated to X-ray fluorescence, and related analytical techniques are discussed. Some examples of trace element analyses in biological materials with synchrotron radiation are presented.

Survey of chemical speciation of trace elements using synchrotron radiation.

Information concerning the chemical state of trace elements in biological systems generally has not been available. Such information for toxic elements and metals in metalloproteins could prove extremely valuable in the elucidation of their metabolism and other biological processes. The shielding of core electrons by binding electrons affect the energy required for creating inner-shell holes. Furthermore, the molecular binding and symmetry of the local environment of an atom affect the absorption spectrum in the neighborhood of the absorption edge. X-ray absorption near-edge structure (XANES) using synchrotron radiation excitation can be used to provide chemical speciation information for trace elements at concentrations as low as 10 ppm. The structure and position of the absorption curve in the region of an edge can yield vital data about the local structure and oxidation state of the trace element in question. Data are most easily interpreted by comparing the observed edge structure and position with those of model compounds of the element covering the entire range of possible oxidation states. Examples of such analyses will be reviewed.