An overview of the new test stand for H⁻ ion sources at FNAL.

A new test stand at Fermi National Accelerator Laboratory (FNAL) is being constructed to carry out experiments to develop and upgrade the present magnetron-type sources of H(-) ions of up to 80 mA at 35 keV in the context of the Proton Improvement Plan. The aim of this plan is to provide high-power proton beams for the experiments at FNAL. The technical details of the construction and layout of this test stand are presented, along with a prospective set of diagnostics to monitor the sources.

A new solid state extractor pulser for the FNAL magnetron ion source.

A new solid state extractor pulser has been installed on the Fermi National Accelerator Laboratory (FNAL) magnetron ion source, replacing a vacuum tube style pulser that was used for over 40 years. The required ion source extraction voltage is 35 kV for injection into the radio frequency quadrupole. At this voltage, the old pulser had a rise time of over 150 µs due to the current limit of the vacuum tube. The new solid state pulsers are capable of 50 kV, 100 A peak current pulses and have a rise time of 9 µs when installed in the operational system. This paper will discuss the pulser design and operational experience to date.

Supercritical water oxidation of a model fecal sludge without the use of a co-fuel.

A continuous supercritical water oxidation reactor was designed and constructed to investigate the conversion of a feces simulant without the use of a co-fuel. The maximum reactor temperature and waste conversion was determined as a function of stoichiometric excess of oxygen in order to determine factor levels for subsequent investigation. 48% oxygen excess showed the highest temperature with full conversion. Factorial analysis was then used to determine the effects of feed concentration, oxygen excess, inlet temperature, and operating pressure on the increase in the temperature of the reacting fluid as well as a newly defined non-dimensional number, NJa representing heat transfer efficiency. Operating pressure and stoichiometric excess oxygen were found to have the most significant impacts on NJa. Feed concentration had a significant impact on fluid temperature increase showing an average difference of 46.4°C between the factorial levels.

Installation and commissioning of the new Fermi National Accelerator Laboratory H- Magnetron.

The Fermi National Accelerator Laboratory (FNAL) 40 year old Cockcroft-Walton 750 keV injectors with slit aperture magnetron ion sources have been replaced with a circular aperture magnetron, Low Energy Beam Transport, Radio Frequency Quadrupole Accelerator, and Medium Energy Beam Transport, as part of the FNAL Proton Improvement Plan. The injector design is based on a similar system at Brookhaven National Laboratory. The installation, commissioning efforts, and source operations to date will be covered in this paper along with plans for additional changes to the original design to improve reliability by reducing extractor spark rates and arc current duty factor.

A genetically engineered Escherichia coli phytase improves nutrient utilization, growth performance, and bone strength of young swine fed diets deficient in available phosphorus.

A 28-d experiment was conducted using 126 crossbred barrows to evaluate the addition of a genetically engineered Escherichia coli phytase to diets that were 0.15% deficient in available P. Growth performance, bone strength, ash weight, and the apparent absorption of P, Ca, Mg, N, energy, DM, Zn, Fe, and Cu were the response criteria. The pigs (2 pigs/pen) averaged 7.61 kg of BW and 30 d of age initially. The low-P basal diet was supplemented with 0, 100, 500, 2,500, or 12,500 units (U) of E. coli phytase/kg of diet, or 500 U of Peniophora lycii phytase/kg of diet. The positive control (PC) diet was adequate in available P. Pigs were fed the diets in meal form. Fecal samples were collected from each pig from d 22 to 27 of the experiment. There were linear and quadratic increases (P < 0.001) in 28-d growth performance (ADFI, ADG, and G:F), bone breaking strength and ash weight, and the apparent absorption (g/d and %) of P, Ca, and Mg (P < or = 0.01 for quadratic) with increasing concentrations of E. coli phytase. Pigs fed the low-P diets containing 2,500 or 12,500 U/kg of E. coli phytase had greater (P < or = 0.01 or P < 0.001, respectively) values for growth performance, bone breaking strength and ash weight, and the apparent absorption (g/d and %) of P, Ca, and Mg than pigs fed the PC diet. The addition of E. coli phytase did not increase the apparent percentage absorption of N, GE, DM, Zn, Fe, or Cu. There were no differences in the efficacy of the E. coli or P. lycii phytase enzymes at 500 U/kg of low-P diet for any criterion measured. In conclusion, there were linear increases in growth performance, bone breaking strength and ash weight, and the apparent absorption of P, Ca, and Mg with increasing addition of E. coli phytase up to 12,500 U/kg of diet. Also, all of these criteria were greater for pigs fed the low-P diets containing 2,500 or 12,500 U of E. coli phytase/kg than for pigs fed the PC diet. The addition of 500, 2,500, or 12,500 U of E. coli phytase/kg of low-P diet reduced P excretion (g/d) in manure by 35, 42, and 61%, respectively, compared with pigs fed the PC diet.

Comparison of growth performance and zinc absorption, retention, and excretion in weanling pigs fed diets supplemented with zinc-polysaccharide or zinc oxide.

Fifty weanling crossbred pigs averaging 6.2 kg of initial BW and 21 d of age were used in a 5-wk experiment to evaluate lower dietary concentrations of an organic source of Zn as a Zn-polysaccharide (Zn-PS) compared with 2,000 ppm of inorganic Zn as ZnO, with growth performance, plasma concentrations of Zn and Cu, and Zn and Cu balance as the criteria. The pigs were fed individually in metabolism crates, and Zn and Cu balance were measured on individual pigs (10 replications per treatment) from d 22 to 26. The basal Phase 1 (d 0 to 14) and Phase 2 (d 14 to 35) diets contained 125 or 100 ppm added Zn as Zn sulfate, respectively, and met all nutrient requirements. Treatments were the basal Phase 1 and 2 diets supplemented with 0, 150, 300, or 450 ppm of Zn as Zn-PS or 2,000 ppm Zn as ZnO. Blood samples were collected from all pigs on d 7, 14, and 28. For pigs fed increasing Zn as Zn-PS, there were no linear or quadratic responses (P > or = 0.16) in ADG, ADFI, or G:F for Phases 1 or 2 or overall. For single degree of freedom treatment comparisons, Phase 1 ADG and G:F were greater (P < or = 0.05) for pigs fed 2,000 ppm Zn as ZnO than for pigs fed the control diet or the diet containing 150 ppm Zn as Zn-PS. For Phase 2 and overall, ADG and G:F for pigs fed the diets containing 300 or 450 ppm of Zn as Zn-PS did not differ (P > or = 0.29) from pigs fed the diet containing ZnO. Pigs fed the diet containing ZnO also had a greater Phase 2 (P < or = 0.10) and overall (P < or = 0.05) ADG and G:F than pigs fed the control diet. There were no differences (P > or = 0.46) in ADFI for any planned comparison. There were linear increases (P < 0.001) in the Zn excreted (mg/d) with increasing dietary Zn-PS. Pigs fed the diet containing ZnO absorbed, retained, and excreted more Zn (P < 0.001) than pigs fed the control diet or any of the diets containing Zn-PS. In conclusion, Phase 2 and overall growth performance by pigs fed diets containing 300 or 450 ppm Zn as Zn-PS did not differ from that of pigs fed 2,000 ppm Zn as ZnO; however, feeding 300 ppm Zn as Zn-PS decreased Zn excretion by 76% compared with feeding 2,000 ppm Zn as ZnO.

Evaluation of various inclusion rates of organic zinc either as polysaccharide or proteinate complex on the growth performance, plasma, and excretion of nursery pigs.

Three experiments were conducted to evaluate the effects of feeding dietary concentrations of organic Zn as a Zn-polysaccharide (Quali Tech Inc., Chaska, MN) or as a Zn-proteinate (Alltech Inc., Nicholasville, KY) on growth performance, plasma concentrations, and excretion in nursery pigs compared with pigs fed 2,000 ppm inorganic Zn as ZnO. Experiments 1 and 2 were growth experiments, and Exp. 3 was a balance experiment, and they used 306, 98, and 20 crossbred pigs, respectively. Initially, pigs averaged 17 d of age and 5.2 kg BW in Exp. 1 and 2, and 31 d of age and 11.2 kg BW in Exp. 3. The basal diets for Exp. 1, 2, and 3 contained 165 ppm supplemental Zn as ZnSO4 (as-fed basis), which was supplied from the premix. In Exp. 1, the Phase 1 (d 1 to 14) basal diet was supplemented with 0, 125, 250, 375, or 500 ppm Zn as Zn-polysaccharide (as-fed basis) or 2,000 ppm Zn as ZnO (as-fed basis). All pigs were then fed the same Phase 2 (d 15 to 28) and Phase 3 (d 29 to 42) diets. In Exp. 2, both the Phase 1 and 2 basal diets were supplemented with 0, 50, 100, 200, 400, or 800 ppm Zn as Zn-proteinate (as-fed basis) or 2,000 ppm Zn as ZnO (as-fed basis). For the 28-d Exp. 3, the Phase 2 basal diet was supplemented with 0, 200, or 400 ppm Zn as Zn-proteinate, or 2,000 ppm Zn as ZnO (as-fed basis). All diets were fed in meal form. In Exp. 1, 2, and 3, pigs were bled on d 14, 28, or 27, respectively, to determine plasma Zn and Cu concentrations. For all three experiments, there were no overall treatment differences in ADG, ADFI, or G:F (P = 0.15, 0.22, and 0.45, respectively). However, during wk 1 of Exp. 1, pigs fed 2,000 ppm Zn as ZnO had greater (P < or = 0.05) ADG and G:F than pigs fed the basal diet. In all experiments, pigs fed a diet containing 2,000 ppm Zn as ZnO had higher plasma Zn concentrations (P < 0.10) than pigs fed the basal diet. In Exp. 1 and 3, pigs fed 2,000 ppm Zn as ZnO had higher fecal Zn concentrations (P < 0.01) than pigs fed the other dietary Zn treatments. In conclusion, organic Zn either as a polysaccharide or a proteinate had no effect on growth performance at lower inclusion rates; however, feeding lower concentrations of organic Zn greatly decreased the amount of Zn excreted.

Copper proteinate in weanling pig diets for enhancing growth performance and reducing fecal copper excretion compared with copper sulfate.

Two 28-d experiments were conducted to evaluate the efficacy of low dietary concentrations of Cu as Cu-proteinate compared with 250 ppm Cu as CuSO4 with growth performance, plasma Cu concentrations, and Cu balance of weanling swine as the criteria. In the production study (Exp. 1), 240 crossbred pigs that averaged 19.8 d of age and 6.31 kg BW initially were group-fed (two or three pigs per pen) the basal diets (Phase 1: d 0 to 14 and Phase 2: d 14 to 28) supplemented with 0 (control), 25, 50, 100, or 200 ppm Cu as Cu-proteinate, or 250 ppm Cu as CuSO4 (as-fed basis). The basal diets contained 16.5 ppm Cu supplied as CuSO4 before supplementation with Cu-proteinate or 250 ppm Cu as CuSO4. There were quadratic responses (P < or = 0.05) in ADFI and ADG for wk 1, Phases 1 and 2, and overall because ADFI was higher for pigs fed 25 or 50 ppm Cu as Cu-proteinate, and ADG increased with increasing Cu-proteinate up to 50 ppm Cu. The Cu-proteinate treatment groups combined had a higher (P < or = 0.05) Phase 2 and overall ADFI and ADG than the CuSO4 group. In the mineral balance study (Exp. 2), 20 crossbred barrows that averaged 35 d of age and 11.2 kg/BW initially were placed in individual metabolism pens with total urine and fecal grab sample collections on d 22 to 26. Treatments were the basal Phase 2 diet supplemented with 0, 50, or 100 ppm Cu as Cu-proteinate, or 250 ppm Cu as CuSO4 (as-fed basis). Treatments did not differ in growth performance criteria. There were linear increases (P < 0.001) in Cu absorption, retention, and excretion (milligrams per day) with increasing Cu-proteinate. Pigs fed 100 ppm Cu as Cu-proteinate absorbed and retained more Cu and excreted less Cu (mg/d, P < or = 0.003) than pigs fed 250 ppm Cu as CuSO4. Plasma Cu concentrations increased linearly (P = 0.06) with increasing Cu-proteinate. In conclusion, weanling pig growth performance was increased by 50 or 100 ppm Cu as Cu-proteinate in our production Exp. 1, but not in our balance Exp. 2, compared with 250 ppm Cu as CuSO4. However, 50 or 100 ppm Cu as Cu-proteinate increased Cu absorption and retention, and decreased Cu excretion 77 and 61%, respectively, compared with 250 ppm Cu as CuSO4.

Low-phytic acid barley improves calcium and phosphorus utilization and growth performance in growing pigs.

Thirty-five crossbred barrows averaging 13.5 kg starting BW were used in a 35-d experiment to compare the availability of P and the nutritional value of two near-isogenic progeny of the barley cultivar 'Harrington'. Low-phytic acid barley (LPB, 0.35% total P, 0.14% phytic acid P) was homozygous for the low-phytic acid 1-1 allele, and the normal barley (NB, 0.35% total P, 0.24% phytic acid P) was homozygous for the normal allele of that gene. Pigs were fed individually twice daily in metabolism pens. Barley was the only source of phytate in semipurified diets, 1 to 3. Diet 1 contained 75% NB, 0.14% estimated available P (aP), and 0.50% Ca. Diet 2 contained 75% LPB, 0.22% aP, and 0.50% Ca. No inorganic P (iP) was added to Diets 1 and 2 in order to measure the animal response to the different concentrations of aP in these cultivars. Diet 3 was NB Diet 1 supplemented with iP to equal the concentration of aP in LPB Diet 2. Practical barley-soybean meal (SBM)-type diets, NB Diet 4 and LPB Diet 5, were formulated to meet all minimum nutrient requirements, and contained 0.30% aP and 0.65% Ca. For the semipurified diets, pigs fed LPB Diet 2 had higher (P < or = 0.05) bone ash weight, bone breaking strength, P absorption and retention, and Ca absorption and retention compared with pigs fed NB Diet 1, with a trend (P = 0.10) for pigs fed LPB Diet 2 to have a higher ADG and gain:feed ratio than pigs fed NB Diet 1. However, pigs fed LPB Diet 2 or NB Diet 3 were not different (P > or = 0.3) in growth performance, fresh bone weight, fat-free dry bone weight, bone ash, bone breaking strength, or N utilization. This indicates that LPB and NB were equal in nutritional value after supplementation of NB with iP to equal the estimated aP in LPB. For the practical barley-SBM diets, there were no differences (P > or = 0.4) between pigs fed NB Diet 4 or LPB Diet 5 for growth performance, fresh bone weight, bone breaking strength, the percentages of P and Ca utilization, or N, DE, and ME utilization. The use of LPB in pig diets reduced P excretion in swine waste by 55% and 16% in our semipurified and practical diets, respectively, compared with NB. Using our in vitro procedure designed to mimic the digestive system of the pig, the availability of P for pigs was estimated at 52% for LPB and 32% for NB.

Effects of dietary calcium:phosphorus ratios on apparent absorption of calcium and phosphorus in the small intestine, cecum, and colon of pigs.

Thirty-two crossbred barrows were used to investigate the effects of dietary Ca:total P (tP) ratios in phytase-supplemented diets on the apparent absorption of P and Ca in the small intestine, cecum, and colon. Three Ca:tP ratio treatments (1.5:1, 1.3:1, or 1.0:1) were created by adjusting the amount of ground limestone added to the basal low-P grower (.39% tP including .07% added inorganic P) and finisher (.32% tP without added inorganic P) diets. All low-P ratio diets were supplemented with Natuphos phytase at 500 units/kg. A positive control diet without phytase supplementation contained adequate P and Ca to meet dietary requirements. At 123 kg, the pigs were slaughtered and the contents of ileum, cecum, and colon were collected. Lowering the dietary Ca:tP ratio in the diets containing phytase linearly increased (P < .01) the apparent absorption (% and g/d) of P in the small intestine, but Ca absorption was not affected. Pigs fed the low-P diet with a Ca:tP ratio of 1.0:1 had an apparent absorption (g/d) of P or Ca similar to that of pigs fed the control diet, which was adequate in Ca and P. Averaged across all diets, the apparent absorption of P was highest when measured at the cecum, and the apparent absorption of Ca was highest when measured at the colon. In conclusion, lowering the dietary Ca:tP ratio to 1.0:1 in a low-P diet containing phytase increased the apparent absorption of P in the small intestine. Furthermore, a significant amount of P was absorbed in the cecum.

Lowering the dietary calcium to total phosphorus ratio increases phosphorus utilization in low-phosphorus corn-soybean meal diets supplemented with microbial phytase for growing-finishing pigs.

Crossbred growing-finishing pigs (n = 120) were used to investigate the effect of three dietary Ca:total P (tP) ratios (1.5:1, 1.3:1, or 1.0:1) on P utilization in low-P corn-soybean meal diets supplemented with microbial phytase at 500 phytase units/kg. The basal grower (23 to 54 kg BW) diet contained .39% tP including .07% added inorganic P (iP), and the basal finisher (54 to 123 kg BW) diet contained .32% tP without added iP. An adequate-P positive control diet without phytase supplementation contained .60% Ca and .50% tP during the growing phase and .50% Ca and .40% tP during the finishing phase. Lowering the Ca:tP ratio linearly increased ADG during the growing phase (P < .03) and overall (P < .08), gain:feed ratio during the growing phase (P < .001), and P absorption during the finishing phase (P < .04). Lowering the Ca:tP ratio linearly increased BW at slaughter (P < .02), carcass weight (P < .04), bone breaking strength (P < .04), and bone ash weight (P < .06), whereas dressing percentage and backfat depth remained unchanged. In conclusion, pig performance and P utilization were increased by lowering the Ca:tP ratio from 1.5:1 to 1.0:1 in low-P corn-soybean meal diets supplemented with microbial phytase.

Soaking increases the efficacy of supplemental microbial phytase in a low-phosphorus corn-soybean meal diet for growing pigs.

Sixty-three crossbred barrows averaging 18.7 kg initial BW were used in a 6-wk study of the effects of soaking on the efficacy of supplemental microbial phytase (Natuphos, BASF) in a low-P corn-soybean meal diet. The basal corn-soybean meal diet contained .06% available P, .32% total P, and .55% Ca with no added inorganic P. The basal diet was supplemented with 0, 250, or 500 phytase units (PU)/kg of diet. The diet was fed dry or soaked (2 parts water:1 part diet and mixed for 2 h at 30 degrees C before feeding) in a 3 x 2 factorial arrangement. A positive control diet was supplemented with inorganic P and provided .23% available P, .48% total P, and .60% Ca. Pigs were individually penned and fed their respective diets to appetite in four equal meals daily. There were no soaking x phytase interactions (P > .1 to .6) for growth performance criteria. Daily gain and gain/feed ratio were increased (P < .01) by soaking and increased linearly (P < .01) by phytase. Daily feed intake was increased linearly (P < .01) by phytase. There were soaking x phytase quadratic interactions (P < .01) for apparent P absorption criteria because soaking the 250 PU/kg diet increased P absorption similar to that obtained with the 500 PU/kg diet fed dry. Apparent P absorption criteria were increased by soaking (P < .01) and were increased linearly (P < .001) and quadratically (P < .03) by phytase. Phytase reduced fecal P excretion 37 to 40% with dry feeding (P < .03) and 48 to 49% with soaking (P < .01).

Vasoactive intestinal peptide stimulates active K+ transport and Na(+)-K(+)-Cl- cotransport in HT-29 cells.

Regulation of active K+ influx and Na(+)-K(+)-Cl- cotransport activity in HT-29 cells by vasoactive intestinal peptide (VIP) was investigated. Both active K+ influx, defined as the ouabain-sensitive component, and Na(+)-K(+)-Cl- cotransport, defined as the ouabain-resistant bumetanide-sensitive component, of total K+ uptake were increased by VIP. VIP increased the maximum velocity (Vmax) values for both components with no change in apparent Michaelis constant (Km) values. Three lines of evidence support the role of adenosine 3',5'-cyclic monophosphate (cAMP) as a mediator of the VIP effects. 1) The rank order potencies of VIP and peptide histidineisoleucineamide (PHI) in binding and cAMP production (J. T. Turner, S. B. Jones, and D. B. Bylund, Peptides Fayetteville 7: 849, 1986) and K+ uptake were consistent; 2) alpha 2-adrenergic agonists inhibited both VIP-stimulated cAMP production (J. T. Turner, C. Ray-Prenger, and D. B. Bylund, Mol. Pharmacol. 28: 422, 1985) and K+ uptake; and 3) forskolin, but not dideoxyforskolin, mimicked the effects of VIP on K+ uptake. Because amiloride blocked the VIP-stimulated active K+ component, the VIP effects on active K+ influx may be secondary to a Na(+)-H+ antiporter-mediated increase in cellular Na+ content. Additional experiments indicated that pretreatment of cells with a protein kinase C activator, previously shown to decrease basal Na(+)-K(+)-Cl- cotransport activity and the apparent number of cotransporters in HT-29 cells (C. C. Franklin, J. T. Turner, and H. D. Kim, J. Biol. Chem. 264: 6667, 1989), did not change the magnitude of response of the remaining cotransporters after adenylate cyclase activation.(ABSTRACT TRUNCATED AT 250 WORDS)

Peritoneal malignant mesothelioma versus serous papillary adenocarcinoma. A histochemical and immunohistochemical comparison.

In order to evaluate adjunctive histologic methods for separating mesothelioma (MM) and serous adenocarcinoma (SC), we studied 28 and 46 respective cases histochemically and immunohistochemically. Ten serous adenocarcinomas arose primarily in extraovarian sites within the abdomen. Diagnoses in each case were established retrospectively by a combination of electron microscopy and clinicopathologic correlation. A panel of antibodies to cytokeratin (CK), epithelial membrane antigen (EMA), B72.3, placental alkaline phosphatase (PLAP), S-100 protein, carcinoembryonic antigen (CEA), Leu M1, CA-125, and amylase (AM) was applied to paraffin sections of each case. Serous carcinoma was reactive for neutral mucins whereas mesothelioma was not; however, only 50% of adenocarcinoma cases stained in this manner. Peritoneal mesothelioma showed reactivity for CK (28 of 28 cases), EMA (24 of 28 cases), AM (five of 28 cases), CA-125 (four of 28 cases), and S-100 protein (three of 28 cases), but lacked B72.3, PLAP, and CEA. Three mesotheliomas expressed Leu M1, but in an extremely focal distribution. Serous carcinoma reacted for CK (46 of 46 cases), EMA (46 of 46 cases), CA-125 (42 of 46 cases), S-100 protein (40 of 46 cases), Leu M1 (34 of 46 cases; with diffuse staining), B72.3 (33 of 46 cases), PLAP (29 of 46 cases), AM (15 of 46 cases), and CEA (six of 46 cases). Two profiles (S-100 + B72.3; S-100 + PLAP) were seen in 41 of 46 serous adenocarcinoma cases but were absent in all mesotheliomas. Hence, these combinations of determinants are effective in separating such neoplasms diagnostically. Moreover, diffuse reactivity for Leu M1, B72.3, PLAP, or CEA in papillary peritoneal neoplasms appears to exclude the possibility of mesothelioma; however, focal Leu M1 reactivity may indeed be seen in mesothelioma. Although CA-125 is a sensitive marker for serous carcinoma, it is not effective in distinguishing it from mesothelioma.

Isolated valvular amyloid.

Two cases are described of grossly evident amyloid infiltration of the cardiac valves, while only minor deposits were found in other locations. The right-sided valves were more heavily involved than the left. No pre-existing disease of the valves was found, and the lesions appeared to have no adverse effect upon the subjects. Only one other similar case was found in the literature. The only consistently associated condition among the recognized cases was advancing age. The name proposed for the condition is isolated valvular amyloid.

Serous papillary carcinomas arising from the peritoneum and ovaries. A clinicopathologic and immunohistochemical comparison.

Primary papillary serous carcinoma arising from the peritoneal surface (serous surface papillary carcinoma; SSPC) is a distinctive neoplasm with a histomorphologic resemblance to serous ovarian papillary carcinoma (SOPC). To determine if these tumors are similar antigenically, we studied 13 examples of SSPC and 31 of SOPC immunohistochemically. Antibodies to several determinants known to occur in the Müllerian epithelium were employed. Both neoplasms were uniformly positive for cytokeratin and epithelial membrane antigen (EMA); in addition, SSPC and SOPC were similar in quantitative and qualitative reactivity for B72.3 antigen, carcinoembryonic antigen, Leu M1, CA-125 antigen, LN1, LN2, MB2, S100 protein, placental alkaline phosphatase, and amylase. Residual nonneoplastic mesothelium failed to express any of these antigens except for cytokeratin, EMA, and CA-125. The clinical behavior of SSPC was similar to that of high-stage SOPC; all patients with adequate follow-up died of their tumors. These results suggest that SSPC and SOPC are analogous lesions, with respect to their cellular differentiation. Moreover, it would appear that both neoplasms display only a limited immunophenotypic homology to the mesothelium.

Vasoactive intestinal peptide receptor/adenylate cyclase system: differences between agonist- and protein kinase C-mediated desensitization and further evidence for receptor internalization.

In this study we have characterized and compared the regulation of the HT29 cell vasoactive intestinal peptide receptor/adenylate cyclase system (VIP-R/AC) by the VIP-R agonist peptide histidineisoleucineamide (PHI) and by activators of protein kinase C (PKC) including phorbol 12-myristate, 13-acetate (PMA) and mezerein. Preincubation with either PHI or PKC activator decreased maximum VIP-stimulated AC activity and decreased the number of cell surface VIP-R. A [125I]VIP binding assay using solubilized VIP-R of the plasma membrane and light vesicle fractions from sucrose density step gradients was developed as a more direct measure of VIP-R internalization. Preincubation with PHI or PMA decreased plasma membrane fraction [125I]VIP binding and increased binding in the light vesicle fraction, thus providing the most direct evidence to date for translocation of VIP-R per se from the plasma membrane to another, presumably intracellular, compartment. Two experimental approaches differentiated between agonist and PKC activator regulation of VIP-R/AC. The protein kinase inhibitors H-7 and staurosporine blocked mezerein-, but not PHI-, induced losses of cell surface VIP-R. Also, down-regulation of PKC did not block PHI-induced loss of cell surface VIP-R. Thus, although both agonist and PKC activators can lead to desensitization and internalization of VIP-R, PKC is apparently not involved in the mechanisms of agonist-induced desensitization.