Electrical current through individual pairs of phosphorus donor atoms and silicon dangling bonds.

Nuclear spins of phosphorus [P] donor atoms in crystalline silicon are among the most coherent qubits found in nature. For their utilization in scalable quantum computers, distinct donor electron wavefunctions must be controlled and probed through electrical coupling by application of either highly localized electric fields or spin-selective currents. Due to the strong modulation of the P-donor wavefunction by the silicon lattice, such electrical coupling requires atomic spatial accuracy. Here, the spatially controlled application of electrical current through individual pairs of phosphorus donor electron states in crystalline silicon and silicon dangling bond states at the crystalline silicon (100) surface is demonstrated using a high-resolution scanning probe microscope operated under ultra-high vacuum and at a temperature of 4.3 K. The observed pairs of electron states display qualitatively reproducible current-voltage characteristics with a monotonous increase and intermediate current plateaus.

Dynamic tunneling force microscopy for characterizing electronic trap states in non-conductive surfaces.

Dynamic tunneling force microscopy (DTFM) is a scanning probe technique for real space mapping and characterization of individual electronic trap states in non-conductive films with atomic scale spatial resolution. The method is based upon the quantum mechanical tunneling of a single electron back and forth between a metallic atomic force microscopy tip and individual trap states in completely non-conducting surface. This single electron shuttling is measured by detecting the electrostatic force induced on the probe tip at the shuttling frequency. In this paper, the physical basis for the DTFM method is unfolded through a physical model and a derivation of the dynamic tunneling signal as a function of several experimental parameters is shown. Experimental data are compared with the theoretical simulations, showing quantitative consistency and verifying the physical model used. The experimental system is described and representative imaging results are shown.

Short communication: Substituting dry distillers grains with solubles and rumen-protected amino acids for soybean meal in late-lactation cows' diets based on corn silage or ryegrass silage.

Excess protein in dairy cattle diets increases production costs and contributes to environmental pollution. The objective of the present study was to evaluate the effect of feeding dry distillers grains with solubles (DDGS) supplemented with rumen-protected Lys and Met in place of solvent-extracted soybean meal on the performance of late-lactation cows. Two experiments were carried out, with each using 24 late-lactating dairy cows distributed among 4 pens. In trial 1, corn silage was the main forage source. Control (HP1) total mixed ration (TMR) contained 16.3% crude protein (CP) with soybean meal as the main protein source. Treatment TMR (LP1) had 13.7% CP when soybean meal was replaced with DDGS and rumen-protected Lys and Met. Forage in trial 2 was ryegrass silage; control TMR (HP2; 15.4% CP) contained soybean meal and rumen-protected Met, whereas treatment TMR (LP2; 13.8% CP) contained DDGS and rumen-protected Lys and Met. Trials were analyzed as crossover design using the MIXED procedure of SAS (SAS Institute Inc., Cary NC) with cow as sampling unit and pen as the experimental unit. Treatments were similar in dry matter intake (21.0 and 20.4 kg/cow per day for HP1 and LP1, respectively) and milk yield (20.7 and 20.5 kg/cow per day for HP1 and LP1, respectively) during trial 1. Milk composition was similar between treatments, averaging 4.22, 3.73, 4.54, and 9.15, respectively, for fat, protein, lactose, and solids nonfat. Milk urea nitrogen decreased from 17.2 mg/dL for HP1 to 9.93 mg/dL for LP1. In trial 2, no significant differences were observed for dry matter intake (21.4 and 20.9 kg/cow per day for HP2 and LP2, respectively), milk yield (28.1 and 26.6 kg/d for HP2 and LP2, respectively), fat yield (0.99 vs. 0.92 kg/d for HP2 and LP2, respectively), protein yield (0.94 vs. 0.86 kg/d for HP2 and LP2, respectively) and lactose yield (1.37 vs. 1.28 for HP2 and LP2, respectively). Milk urea nitrogen decreased from 9.88 mg/dL with HP2 to 6.39 mg/dL with the LP2 treatment. Milk N efficiency tended to be higher for LP treatments in trial 1, but not in trial 2. Low milk urea N suggested nitrogen losses to the environment may be lower when cows were fed diets based on DDGS in both trials. The studies indicated that DDGS with rumen-protected Lys and Met could substitute solvent-extracted soybean meal in low-protein corn silage- and ryegrass silage-based diets for late-lactation dairy cows averaging 20.6 or 27.4 kg of milk/d, respectively.

Short communication: Effects of increasing protein and energy in the milk replacer with or without direct-fed microbial supplementation on growth and performance of preweaned Holstein calves.

Forty-four Holstein calves were fed a direct-fed microbial (DFM) and 1 of 2 milk replacers to evaluate calf performance and growth. Treatments were (1) a control milk replacer [22:20; 22% crude protein (CP) and 20% fat], (2) an accelerated milk replacer (27:10; 27% CP and 10% fat), (3) the control milk replacer with added DFM (22:20+D), and (4) the accelerated milk replacer with added DFM (27:10+D). Dry matter intake, rectal temperatures, respiration scores and rates, and fecal scores were collected daily. Body weight, hip and withers height, heart girth, blood, and rumen fluid samples were collected weekly. Effects of treatment, sex, week, and their interactions were analyzed. Calves fed an accelerated milk replacer, regardless of DFM supplementation, consumed more CP and metabolizable energy in the milk replacer. No treatment differences were found for starter intake or intake of neutral detergent fiber or acid detergent fiber in the starter. Calves fed the accelerated milk replacer had greater preweaning and weaning body weight compared with calves fed the control milk replacer. Average daily gain was greater during the preweaning period for calves fed the accelerated milk replacer, but the same pattern did not hold true during the postweaning period. Feed efficiency did not differ among treatments. Hip height tended to be and withers height and heart girth were greater at weaning for calves fed the accelerated milk replacer compared with calves fed the control milk replacer. Fecal scores were greatest in calves fed DFM. Overall acetate, propionate, butyrate, and n-valerate concentrations were lower in calves fed the accelerated milk replacer, but DFM did not have an effect. Rumen pH was not different. Blood metabolites were unaffected by DFM supplementation, but calves fed the accelerated milk replacer had increased partial pressure of CO2, bicarbonate, and total bicarbonate in the blood. Direct-fed microbial supplementation did not appear to benefit the calf in this trial.

Stable inhibition of specific estrogen receptor α (ERα) phosphorylation confers increased growth, migration/invasion, and disruption of estradiol signaling in MCF-7 breast cancer cells.

Elevated phosphorylation of estrogen receptor α (ERα) at serines 118 (S118) and 167 (S167) is associated with favorable outcome for tamoxifen adjuvant therapy and may serve as surrogate markers for a functional ERα signaling pathway in breast cancer. It is possible that loss of phosphorylation at S118 and/or S167 could disrupt ERα signaling, resulting in aggressive ERα-independent breast cancer cells. To this end, MCF-7 breast cancer cells were stably transfected with an ERα-specific short hairpin RNA that reduced endogenous ERα. The resulting cell line was stably transfected with wild-type ERα (ER-AB cells), or ERα containing serine to alanine mutation at S118 or S167 (S118A cells and S167A cells, respectively). These stable cell lines expressed approximately equivalent ERα compared with parental MCF-7 cells and were evaluated for growth, morphology, migration/invasion, and ERα-regulated gene expression. S118A cells and S167A cells exhibited increased growth and migration/invasion in vitro. Forward- and side-scatter flow cytometry revealed that S167A cells were smaller in size, and both S118A and S167A cells exhibited less cellular complexity. S118A and S167A cells expressed pancytokeratin and membrane localization of ß-catenin and did not express vimentin, indicating retention of epithelial lineage markers. Expression of ERα-target genes and other genes regulated by ERα signaling or involved in breast cancer were markedly altered in both S118A and S167A cells. In summary, attenuated phosphorylation of ERα at S118 and S167 significantly affected cellular physiology and behavior in MCF-7 breast cancer cells, resulting in increased growth, migration/invasion, compromised expression of ERα target genes, and markedly altered gene expression patterns.

Electronic characterization of individual monolayer protected Au clusters by single electron tunneling force spectroscopy.

Gold monolayer protected clusters (MPCs) exhibit strong quantum confinement effects and size dependent electronic, optical and chemical properties. Chemical tuning of these properties can be achieved by established synthesis methods, providing an excellent system for the study of the relationship between chemical and electronic structure. In this paper, the first electronic spectra of individual Au MPCs (Au(25)) acquired by single electron tunneling force spectroscopy on non-conducting silicon dioxide surfaces are reported. A HOMO-LUMO (highest occupied molecular orbital-lowest unoccupied molecular orbital) energy gap is observed in the Au(25) spectra. Hysteretic charging of the particles is also observed while obtaining the energy spectra. The new single electron tunneling measurement methodology is described. A model explaining the measurements supports the existence of mid-HOMO-LUMO gap defect states.

Influence of calcium and phosphorus feeding on markers of bone metabolism in transition cows.

A study was carried out to verify the effect of Ca and P levels on production, digestibility, and serum bone metabolism biomarkers in dairy cows. Fifty-two nonlactating multiparous cows (>or=3 lactations) were confined in a free-stall barn approximately 20 d before calving. A standard close-up diet was fed to cows once daily until d 2 postpartum. Cows were randomly assigned to 1 of 4 dietary treatments arranged in a 2 x 2 factorial approach averaging 0.64% Ca for high Ca (HCa), 0.46% Ca for low Ca (LCa), 0.47% P for high P (HP), and 0.38% P for low P (LP) on a dry matter basis. Experimental diets were fed twice daily from 3 d in milk (DIM) until 31 DIM. Intake and milk yield were recorded daily. Milk samples were collected on d 28, 29, and 30 postpartum for components analyses. Blood samples were drawn 10 d before expected calving, at calving, and at 15 and 30 DIM for serum analyses of osteocalcin, a biomarker of bone accretion, and pyridinoline, a biomarker of bone resorption. Total fecal collection was conducted when cows in a block averaged 20 DIM. Intake and production traits were not significantly affected by any of the dietary treatments. Cows averaged nearly 21 kg/d dry matter intake and 44 kg/d milk yield from 6 to 31 DIM. There were no significant differences across treatments in body weight or body condition score loss. Phosphorus intake, P fecal output, P digestibility, and P apparent absorption were affected by dietary P content. Calcium intake was higher with HCa, but Ca fecal output, digestibility, and apparent absorption showed an interaction between dietary Ca and dietary P. Calcium fecal output was 100.6 g/d for cows fed HCaHP, intermediate for cows on the HCaLP diet (89 g/d), and similar among cows fed the 2 LCa diets (70 g/d with LCaHP and 75 with LCaLP). There was no significant effect of Ca or P on osteocalcin measurements. Pyridinoline concentrations were affected by dietary Ca levels and tended to have a significant dietary Ca x dietary P interaction. Phosphorus apparent digestibility occurred independently of dietary Ca levels. Results of this study suggest that more bone was mobilized in cows fed LCa diets, but excess dietary P caused greater and prolonged bone mobilization regardless of dietary Ca content.

Atomic scale imaging and spectroscopy of individual electron trap states using force detected dynamic tunnelling.

We report the first atomic scale imaging and spectroscopic measurements of electron trap states in completely non-conducting surfaces by dynamic tunnelling force microscopy/spectroscopy. Single electrons are dynamically shuttled to/from individual states in thick films of hafnium silicate and silicon dioxide. The new method opens up surfaces that are inaccessible to the scanning tunnelling microscope for imaging and spectroscopy on an atomic scale.

The influence of dietary selenium on common indicators of selenium status and liver glutathione peroxidase-1 messenger ribonucleic acid.

The objective of this research was to determine the influence of dietary Se on various indicators of Se status and relative liver glutathione peroxidase 1 (GPx-1) messenger RNA (mRNA) levels in growing Holstein bull calves. Calves (n = 14, 7/diet) were started 28 d after birth on a Se-adequate (SeA) or Se-deficient diet (SeD) and maintained on the diet until 180 d of age. Blood samples were taken from each calf for determination of erythrocyte GPx-1 and plasma GPx-3 activities and plasma Se concentration on d 28 of age, every 28 d thereafter, and at 180 d of age. To assess liver Se and GPx-1 mRNA, 3 calves were first killed at d 21 of age for baseline (BSL) measurements, and 4 calves from each treatment were killed at trial conclusion. Feed intake and ADG were not affected (P = 0.62) by dietary Se concentrations. However, liver Se concentration was greater (P < 0.05) for BSL calves and SeA calves than SeD calves, but no difference (P = 0.68) was observed between BSL calves and SeA calves. Plasma Se was greater for SeA calves (P < 0.01) than for SeD calves by d 56 of age. The GPx-1 activity was greater in SeA calves (P < 0.01) by d 84 of age, whereas GPx-3 activity was greater in SeA calves, but not until d 180 of age (P < 0.01). There was a 50% decrease in GPx-1 mRNA for the SeD calves (P < 0.05) compared with SeA calves. Thus, relative GPx-1 mRNA transcript level is reflective of Se status in the bovine. Furthermore, 152 d on a semi-purified, SeD diet is adequate to create a Se deficiency in growing Holstein bull calves started on a SeD diet at 28 d of age.

Effects of methionine and lysine on fermentation in vitro and in vivo, nutrient flow to the intestine, and milk production.

Three experiments were conducted to investigate the feasibility of using crystalline methionine and lysine as protein supplements for lactating Holstein cows. In the first experiment, Met (dl-methionine) and Lys (l-lysine-HCl) were added to diets used in continuous culture bioreactors to estimate optimal concentrations for use in subsequent in vivo experiments. The second experiment measured ruminal fermentation and nutrient flow to the small intestine when Met and Lys were top-dressed on diets fed to nonlactating cows. The third experiment measured lactation performance when Met and Lys were added to diets fed to late-lactation cows. Providing 0.29 and 2.27% of dry matter as Met and Lys, respectively, provided the largest improvement in fermentation in vitro and these concentrations were used in subsequent experiments. When Met and Lys were top-dressed on diets fed to nonlactating cows, no changes in total tract N digestion were observed. No changes in microbial protein production or ruminal fermentation were observed. Adding Met and Lys did not change production or efficiency of production of milk or milk components by late lactation cows. These data indicate that providing supplemental Met and Lys during late lactation does not significantly improve the protein status of the cow and therefore may not improve milk production.

Effects of zinc and sodium monensin on ruminal degradation of lysine-HCl and liquid 2-hydroxy-4-methylthiobutanoic acid.

Four nonlactating, mature, Holstein cows were fitted with ruminal cannula and used in a 4 x 4 Latin square-designed experiment to evaluate the impact of supplemental Zn and monensin on ruminal degradation of Lys and liquid 2-hydroxy-4-methylthiobutanoic acid (HMB). Cows were fed 4.54 kg (as fed) of alfalfa hay top-dressed with 4.54 kg (as fed) concentrate once daily. Concentrates were formulated to provide 0 or 500 mg/kg of Zn as ZnSO4 and 0 or 40 mg/kg of monensin in the total diet. Zinc supplementation provided approximately 22-fold greater dietary Zn than estimated by NRC requirements. On d 14 of each period, cows were dosed via the rumen cannula with 50 g of HMB and 100 g of Lys-HCl, and the concentrations of Lys and HMB were monitored every 0.5 h for 8 h. Supplemental Zn tended to decrease the proportion of acetate in ruminal fluid postfeeding and increased the proportion of propionate in ruminal fluid postfeeding. Supplemental Zn increased mean fluid passage rate from the rumen. Monensin decreased the proportion of acetate and increased the mean proportion of propionate in ruminal fluid, resulting in a decrease in the ratio of acetate to propionate. Monensin also increased the mean fluid passage rate from the rumen. Neither Zn nor monensin affected the apparent rate of ruminal disappearance of HMB or Lys. However, Zn and monensin interacted to alter the ruminal degradability of free Lys but not HMB. These data indicate that Zn and monensin may interact to alter ruminal degradability of free amino acids.

Flow paths of plant tissue residues and digesta through gastrointestinal segments in Spanish goats and methodological considerations.

A sequence of eight twice-daily meals, each marked with different rare earth elements, was fed to 24 Spanish goats (BW = 20.6 +/- 1.94 kg) to produce meal-based profiles of rare earth markers within segments of the gastrointestinal digesta on subsequent slaughter. Accumulative mean residence time and time delay of rare earths and segmental and accumulative mean residence times of indigestible NDF (IDF) were estimated for each sampled segment. Diets consisted of ad libitum access to bermudagrass hay with a limit feeding of one of four supplements: 1) minerals (basal, B); 2) B + energy (E); 3) B + CP (CP); or 4) B + E + CP for 84 d. Mean daily intake (g/kg of BW) during the 5 d before slaughter differed (P < 0.05) via diet for DM but not for IDF (8.0 +/- 0.35 g/kg of BW). Larger estimates of cumulative mean residence time for IDF vs. rare earths were suggested to be the consequence of a meal-induced bias in the single measurement of IDF pool size by anatomical site. The rare earth compartment method was considered more reliable than the IDF pool dilution method because it yielded flow estimates based on the flux of eight meal-dosed rare earth markers over 4 d and was independent of anatomical definitions of pool size. Statistically indistinguishable estimates for gastrointestinal mean residence times for IDF and rare earths conform to assumed indelibility for the specifically applied rare earths and indigestibility of IDF. The potentially digestible NDF (PDF):IDF ratio of dietary fragments (0.8) progressively decreased in the following order: caudodorsal reticulorumen (0.390) > crainodorsal reticulorumen (0.357) approximately reticulum (0.354) > mid-dorsal reticulorumen (0.291) approximately ventral reticulorumen (0.286), to that within the omasal folds and in the abomasum (0.259). Such a gradient of progressively aging mixture of plant tissue fragments is consistent with age-dependent flow paths established in the reticulorumen and flowing to the omasum and abomasum. Such heterogeneity of fragment ages within the reticulorumen is also indicated by the superior fit of marker dose site double dagger marker sampling site model assumptions. Additionally, cyclic meal- and rumination-induced variations in escape rate occur. Estimates of mean escape rates over days, needed for the practice of ruminant nutrition, must consider the complex interactions among plant tissues and the dynamics of their ruminal digestion of PDF.

Thyrotropin releasing hormone interactions with growth hormone secretion in horses.

Light horse mares, stallions, and geldings were used to 1) extend our observations on the thyrotropin releasing hormone (TRH) inhibition of GH secretion in response to physiologic stimuli and 2) test the hypothesis that stimulation of endogenous TRH would decrease the normal rate of GH secretion. In Exp. 1 and 2, pretreatment of mares with TRH (10 microg/kg BW) decreased (P < 0.001) the GH response to exercise and aspartate infusion. Time analysis in Exp. 3 indicated that the TRH inhibition lasted at least 60 min but was absent by 120 min. Administration of a single injection of TRH to stallions in Exp. 4 increased (P < 0.001) prolactin concentrations as expected but had no effect (P > 0.10) on GH concentrations. Similarly, 11 hourly injections of TRH administered to geldings in Exp. 5 did not alter (P > 0.10) GH concentrations either during the injections or for the next 14 h. In Exp. 5, it was noted that the prolactin and thyroid-stimulating hormone responses to TRH were great (P < 0.001) for the first injection, but subsequent injections had little to no stimulatory effect. Thus, Exp. 6 was designed to determine whether the inhibitory effect of TRH also waned after multiple injections. Geldings pretreated with five hourly injections of TRH had an exercise-induced GH response identical to that of control geldings, indicating that the inhibitory effect was absent after five TRH injections. Retrospective analysis of pooled, selected data from Exp. 4, 5, and 6 indicated that endogenous GH concentrations were in fact lower (P < 0.01) from 45 to 75 min after TRH injection but not thereafter. In Exp. 7, 6-n-propyl-2-thiouracil was fed to stallions to reduce thyroid activity and hence thyroid hormone feedback, potentially increasing endogenous TRH secretion. Treated stallions had decreased (P < 0.01) concentrations of thyroxine and elevated (P < 0.01) concentrations of thyroid-stimulating hormone by d 52 of feeding, but plasma concentrations of GH and prolactin were unaffected (P > 0.10). In contrast, the GH response to aspartate and the prolactin response to sulpiride were greater (P < 0.05) in treated stallions than in controls. In summary, TRH inhibited exercise- and aspartate-induced GH secretion. The duration of the inhibition was at least 1 h but less than 2 h, and it waned with multiple injections. There is likely a TRH inhibition of endogenous GH episodes as well. Reduced thyroid feedback on the hypothalamic-pituitary axis did not alter basal GH and prolactin secretion.

Effects of feeding milk replacer once versus twice daily on glucose metabolism in Holstein and Jersey calves.

Eighteen Holstein (experiment 1) and 15 Jersey (experiment 2) heifer calves were fed milk replacer once or twice daily to determine effects of feeding frequency on weight gain, starter intake, and glucose metabolism. Body weights were measured weekly from birth to 8 wk. Blood samples were collected at wk 1 through 6 from all calves before and at 30, 60, 90, 120 and 180 min after the morning feeding. Plasma was analyzed for glucose, insulin, glucagon, and nonesterified fatty acids (NEFA). Urine was collected 90 min postfeeding to measure glucose concentration. Treatment did not affect mean starter intake or body weight. In experiments 1 and 2 mean plasma glucagon, glucose, NEFA, and insulin and urinary glucose concentrations were not affected by treatment. There was an interaction of sampling time and treatment for plasma insulin concentrations but not for glucose concentrations in both experiments. Following feeding, calves fed milk replacer once daily had higher insulin concentrations than those fed twice daily. There was an interaction of sampling time and treatment for plasma NEFA concentrations in Jersey calves only. Jersey calves fed milk replacer once daily had higher plasma NEFA concentrations before the morning milk replacer feeding. At wk 3 and 6, frequently sampled intravenous glucose tolerance tests were performed to assess glucose effectiveness, insulin sensitivity, and acute insulin response. In experiments 1 and 2 glucose effectiveness and insulin sensitivity were similar regardless of milk replacer feeding frequency. In Holstein and Jersey calves fed milk replacer twice daily, acute insulin response was greater than in calves fed once daily. However, insulin sensitivity decreased with age, while acute insulin response increased with age. These data suggest that feeding calves milk replacer once daily did not deleteriously affect performance or glucose metabolism regardless of breed.

Increasing access and building equity into mental health services: an examination of the potential for change.

This article explores the use of mental health care services by ethnoracial people in Canada and distinguishes between the reasons for underutilization of services by ethnoracial groups and the barriers which prevent ethnoracial groups from accessing services. Research focusing on Canadian race relations is reviewed to reveal how they are paralleled in the functioning of mainstream mental health care organizations. Existing policies and attitudes are then considered in relation to how they support or impede interventions to increase accessibility to services. Finally, frameworks for organizational change based on multiculturalism and anti-racism are presented, and the advantages and disadvantages of both are articulated.

Is retrieval-induced forgetting an inhibitory process?

Three experiments investigated memory performance in the retrieval practice paradigm (Anderson, Bjork, & Bjork, 1994; Anderson & Spellman, 1995). This paradigm produces a retrieval-induced forgetting effect, wherein practicing some members of a studied category decreases the recall of other members of that category relative to a baseline. Our findings indicate that the retrieval-induced forgetting effect is replicable but that previous findings supporting an inhibitory account of this phenomenon may not be.

A limit on retrieval-induced forgetting.

Retrieving some members of a memory set impairs later recall of semantically related but not unrelated members (M. C. Anderson, R. A. Bjork, & E. L. Bjork, 1994; M. C. Anderson & B. A. Spellman, 1995). The authors investigated whether this retrieval-induced forgetting effect would generalize to testing procedures other than category-cued recall. Although the authors demonstrated a retrieval-induced forgetting effect using a category-cued recall task, they failed to show retrieval-induced forgetting on several different memory tests that used item-specific cues, including a category-plus-stem-cued recall test, a category-plus-fragment-cued recall test, a fragment-cued recall test, and a fragment completion task.

Optical dipole model for photodetection in the near field.

Near-field photodetection optical microscopy (NPOM) is a scanning probe technique that has been developed to perform nanometer-scale optical intensity mapping and spectroscopy. In NPOM a nanometer-scale photodiode detector absorbs power directly as it is scanned in the near field of an illuminated sample surface. A model of photodetection in the near and intermediate fields is presented. A brief review of far-field absorption is given for comparison. Far-field absorption measurements measure the imaginary part of the polarizability to first order. In contrast, photodetection in the near field measures the real part of the polarizability. Other aspects of near-field photodetection are also examined, including contrast mechanisms and lateral resolution. NPOM measurements performed on isolated 300-nm spheres show good agreement with the theory.

Dexamethasone increases the expression of membrane macrophage colony stimulating factor from retrovirally transduced tumor cells expressing macrophage colony stimulating factor.

Many different tumor cell types (breast, ovarian, glioma, liver and colon) were retrovirally transduced with the human macrophage colony stimulating factor (M-CSF) gene (either the membrane associated form [mM-CSF] or the secreted form [sM-CSF]). These cells were tested for their ability to display increased amounts of mM-CSF in response to dexamethasone. M-CSF-transfected tumor cells expressed additional mM-CSF in response to 18-72 h incubations with 3-15 microg/ml dexamethasone, while non-transfected parental cells were unaffected by this treatment. Increased mM-CSF protein expression on the M-CSF transduced cells was observed by flow cytometry and Western blotting using M-CSF specific antibodies. Northern blot analysis revealed an increase in the mM-CSF specific transcripts within the dexamethasone-treated mM-CSF transduced cells, but this was not seen within the non-transfected tumor cells that were treated with dexamethasone. ICAM-1 expression was unaffected by dexamethasone treatment, indicating that this response is mM-CSF specific. All trans-retinal and 1,25-dihydroxy vitamin D3 compounds that have been reported to induce M-CSF expression failed to increase mM-CSF. When dexamethasone-treated mM-CSF transfected clones were used as target cells for macrophage-mediated cytotoxicity assays, an increased killing with the dexamethasone-treated cells was seen. The macrophage-mediated cytotoxicity of these mM-CSF expressing tumor cells was blocked with excess recombinant M-CSF by saturating M-CSF receptors on the macrophage that is required for this form of tumor cell killing. This work suggests the possibility that dexamethasone may prove useful for vaccination purposes using mM-CSF retrovirally transfected tumor cells.

Membrane macrophage colony-stimulating factor on MADB106 breast cancer cells does not activate cytotoxic macrophages but immunizes rats against breast cancer.

Weakly immunogenic, but highly malignant, rat MADB106 breast cancer cells were retrovirally transduced with the membrane form of macrophage colony-stimulating factor (mM-CSF). The cloned mM-CSF-transfected MADB106 cells physically conjugated with macrophages, but were not killed by the macrophages in 48-h cytotoxicity assays. Macrophages killed the mM-CSF-expressing tumors in the presence of noncytotoxic doses of either taxol or taxol plus cisplatin. This indicated that macrophages bind to the mM-CSF expressed on the tumor cells, but for successful macrophage cytotoxicity to occur against mM-CSF-transduced tumor cells other factors must be present. The mM-CSF-transfected tumor cells were rejected when inoculated subcutaneously into normal rats. Cloned MADB106 tumor cells which expressed high amount of mM-CSF were rejected, while tumor cells that displayed lower levels of mM-CSF grew in 60% of the inoculated rats. The mM-CSF-transfected tumors that grew were smaller and had a greater amount of necrosis, compared to the viral vector tumors. Rats that spontaneously rejected the mM-CSF-transfected MADB106 cells showed rechallenge resistance to unmodified parental MADB106 and R3230Ac breast cancers, but not to the F98 glioma. These observations suggest that breast cancer-specific immunity was induced by the inoculation of mM-CSF-expressing MADB106 tumor cells.