Effects of post-mortem aging time and type of aging on palatability of low marbled beef loins.

The study objective was to evaluate the effect of post-mortem aging period (14 to 49days), dry vs. wet (D vs W) type of aging on the palatability of bone-in (BI) beef short loins (n=96) and boneless (BL) strip loins (n=96) possessing United States Department of Agriculture marbling scores between Slight and Small. Warner-Bratzler shear force (WBSF) scores decreased linearly over time (P=0.0001). WBSF was not influenced by aging method or loin type. Aged flavor was higher for DBL than for DBI with WBL and WBI intermediate. Dry aging strip loins increase aged flavor yet did not improve beefy flavor compared to wet aging. Based on objective data and panelist's scores for tenderness, juiciness and aged flavor, a boneless, 28days wet aged strip steak, cooked to 71°C would provide the best combination of eating satisfaction and value.

Consumer evaluation of palatability characteristics of a beef value-added cut compared to common retail cuts.

The objectives of this study were to educate consumers about value-added beef cuts and evaluate their palatability responses of a value cut and three traditional cuts. Three hundred and twenty-two individuals participated in the beef value cut education seminar series presented by trained beef industry educators. Seminar participants evaluated tenderness, juiciness, flavor, and overall like of four samples, bottom round, top sirloin, ribeye, and a value cut (Delmonico or Denver), on a 9-point scale. The ribeye and the value cut were found to be similar in all four attributes and differed from the top sirloin and bottom round. Correlations and regression analysis found that flavor was the largest influencing factor for overall like for the ribeye, value cut, and top sirloin. The value cut is comparable to the ribeye and can be a less expensive replacement.

Effectiveness of oxygen barrier oven bags in low temperature cooking on reduction of warmed-over flavor in beef roasts.

A 3×3×2 factorial was utilized to determine if roast size (small, medium, large), cooking method (open-pan, oven bag, vacuum bag), and heating process (fresh, reheated) prevented warmed-over flavor (WOF) in beef clod roasts. Fresh vacuum bag and reheated open-pan roasts had higher cardboardy flavor scores compared with fresh open-pan roast scores. Reheated roasts in oven and vacuum bags did not differ from fresh roasts for cardboardy flavor. Brothy and fat intensity were increased in reheated roasts in oven and vacuum bags compared with fresh roasts in oven and vacuum bags. Differences in TBARS were found in the interaction of heating process and roast size with the fresh and reheated large, and reheated medium roasts having the lowest values. Based on TBARS data, to prevent WOF in reheated beef roasts, a larger size roast in a cooking bag is the most effective method.

Influence of level of dried distillers grains with solubles on feedlot performance, carcass characteristics, serum testosterone concentrations, and spermatozoa motility and concentration of growing rams.

The objective of this study was to evaluate the effects of dried distillers grains with solubles (DDGS) on ram lamb feedlot performance, carcass characteristics, serum testosterone concentration, and semen quality. One hundred twenty ram lambs (40.4 ± 9.1 kg; Suffolk × western white face) were used in a completely randomized design to determine the effects of DDGS on feedlot performance and carcass characteristics. Rams were allotted into one of three dietary treatments (n = 4 pens/treatment; 10 rams/pen): 1) 0DDGS: 85% corn and 15% commercial market lamb pellet, 2) 15DDGS: 15% DDGS substituted for corn (DM basis), and 3) 30DDGS: 30% DDGS substituted for corn (DM basis). Rams were weighed on consecutive days at the beginning (d 0 and 1) and end (d 96 and 97 and d 116 and 117) of the trial. Scrotal circumference was measured on all rams on d 84, 96, and 116. Semen and blood samples were collected on a subset of 48 rams (4 rams/pen; 16 rams/treatment; n = 4) to evaluate semen quality. Blood samples were collected every 14 d throughout the study. Semen samples were collected on d 84, 98, and 112. Rams were fed to market weight, shipped to a commercial abattoir, and harvested for carcass data collection. Initial BW, final BW, change in scrotal circumference, days on feed, carcass characteristics, serum testosterone concentrations, and spermatozoa motility score were not different (P ≥ 0.23) due to dietary treatment. However, DMI increased linearly (P < 0.001) as DDGS increased in the ration, resulting in a linear increase (P = 0.02) in ADG. Additionally, spermatozoa concentration decreased linearly (P = 0.05) as DDGS concentration increased in the ration. Increasing DDGS in the diet did not have a negative impact on ram feedlot performance or carcass characteristics; however, spermatozoa production may have been negatively affected, necessitating the need for additional research on the impact of DDGS on ram development.

Effects of increasing dosages of zeranol implants on lamb growth, carcass characteristics, blood hormones, and nitrogen metabolism.

The objective of this research was to compare the growth performance, incidence of prolapse and mortality, carcass characteristics, blood hormone concentration, and N balance of lambs implanted with increasing dosages of zeranol. One hundred forty-four crossbred lambs (29.6 ± 4.9 kg) were used in a completely random design and placed into 16 feedlot pens (4 pens/treatment) for a 116 d finishing study. Lambs were fed an 84.7% corn and 15.3% market lamb pellet (DM basis) diet ad libitum. Treatments were 0, 12, 24, and 36 mg zeranol (Ralgro; Schering-Plough), and lambs were implanted in the ear according to treatment on d 0. Lambs were weighed. Thirty lambs (67.6 ± 3.4 kg) and 96 lambs (65.8 ± 5.1 kg) were harvested on d 84 and d 118, respectively. Carcass data were collected 24 h after chill. Blood samples were collected on d 0, 28, 56, 70, 82, 99, and 116 from 64 lambs (29.6 ± 2.1 kg) in the feedlot study (subsample of 4 lambs per pen) and analyzed for thyroxine, triiodothyronine, and IGF-I. A second study was conducted to compare effects of 0, 12, 24, or 36 mg zeranol on N balance in 16 crossbred lambs (34.8 ± 2.1 kg). There were no differences among treatments for BW, ADG, DMI, and G:F (P > 0.05) in the feedlot study. However, there was a linear increase for incidence of prolapse (P = 0.006; 2.78, 5.55, 24.98, and 27.75%, respectively) and mortality (P = 0.005; 0.00, 5.55, 11.10, and 13.88%, respectively) as zeranol dosage increased. Carcass characteristics, blood hormone concentrations, and N balance were not affected by treatment (P > 0.05). These results indicate zeranol increases incidence of prolapse and mortality without increasing growth performance.

Interaction of corn processing and distillers dried grains with solubles on health and performance of steers.

Feeding increased concentrations of distillers dried grains with solubles (DDGS) to ruminants has been avoided due to risks of S toxicity and concerns about animal performance. The objective of this study was to evaluate the influence of feeding an increasing concentration of DDGS and corn processing method on animal performance, incidence of polioencephalomalacia (PEM), and concentration of H(2)S gas in feedlot steers. Sixty steer calves (336 ± 13.2 kg) were individually fed for an average of 136 d in a completely random design with a 3 × 2 factorial arrangement of treatments. Main effects included concentration of DDGS (20, 40, or 60% DM basis) and corn processing method [high-moisture (HMC; 71.7% DM) vs. dry-rolled corn (DRC; 86.2% DM)] resulting in treatments of 1) 20% DDGS with DRC, 2) 40% DDGS with DRC, 3) 60% DDGS with DRC, 4) 20% DDGS with HMC, 5) 40% DDGS with HMC, and 6) 60% DDGS with HMC. Ruminal H(2)S gas concentrations were measured on d 0, 7, 14, 21, 28, 35, 49, 63, and 91 via rumen puncture. Animal performance and carcass characteristic data were collected. The day × corn processing × DDGS interaction for H(2)S gas concentrations was not significant (P = 0.91). Ruminal H(2)S concentration increased with increasing DDGS concentration (P < 0.001) and day (P < 0.001), but was not influenced by corn processing method (P = 0.94). Carcass-adjusted final BW decreased linearly (P = 0.009), whereas carcass-adjusted ADG decreased quadratically (P = 0.05) with increasing concentration of DDGS in the diet. Carcass-adjusted G:F was not affected (P ≥ 0.28) by increasing concentration of DDGS in the diet. Carcass characteristics reflected the decrease in final BW with decreased HCW (P = 0.009), as well as decreased fat depth (P = 0.005) with increasing concentrations of DDGS. The combination of decreased HCW and backfat thickness resulted in decreased (P = 0.02) yield grade with increasing DDGS inclusion. There were no confirmed cases of PEM. In conclusion, corn processing did not influence animal performance, incidence of PEM, or H(2)S concentrations under the conditions of this study. Feeding 60% DDGS in beef cattle finishing diets is not recommended due to poor animal performance.

Effect of thiamine concentration on animal health, feedlot performance, carcass characteristics, and ruminal hydrogen sulfide concentrations in lambs fed diets based on 60% distillers dried grains plus solubles.

Limited data are available regarding the influence of thiamine supplementation on the incidence of polioencephalomalacia (PEM) in lambs fed diets containing increased concentrations of S in the diet (>0.7%). Therefore, our objective was to evaluate the influence of thiamine supplementation on feedlot performance, carcass quality, ruminal hydrogen sulfide gas concentrations, and incidence of PEM in lambs fed a finishing diet containing 60% distillers dried grains with solubles (DDGS; DM basis). Two studies were conducted using completely randomized designs to evaluate the influence of concentration of thiamine supplementation. Study 1 used 240 lambs fed in 16 pens, whereas study 2 used 55 individually fed lambs. Lamb finishing diets contained 60% DDGS, which resulted in a dietary S concentration of 0.73% (DM basis). Treatments diets were based on the amount of supplemental thiamine provided: 1) no supplemental thiamine (CON), 2) 50 mg/animal per day (LO), 3) 100 mg/animal per day (MED), or 4) 150 mg/animal per day (HI). Additionally, in study 2, a fifth treatment was included, which contained 0.87% S (DM basis; increased S provided by addition of dilute sulfuric acid) and provided 150 mg of thiamine/animal per day (HI+S). In study 1, ADG decreased quadratically (P = 0.04), with lambs fed the CON, LO, and MED diets gaining BW at a greater rate than lambs fed the HI diet. In study 1, DMI responded quadratically (P < 0.01), whereas G:F tended to differ linearly (P = 0.08) to concentration of thiamine supplementation, with MED lambs having greater DMI and decreased G:F. No differences (P > or = 0.17) in lamb performance were observed in study 2. In both studies, most carcass characteristics were unaffected, with the exception of a tendency for decreased carcass conformation (study 1; P = 0.09) and greater flank streaking (study 2; P = 0.03). No differences in ruminal hydrogen sulfide concentration (P > 0.05) among treatments were apparent until d 10, at which point lambs fed the LO diet had less hydrogen sulfide concentrations than all other treatments. Lambs fed HI had the greatest concentrations of hydrogen sulfide on d 31 (1.07 g of hydrogen sulfide /m(3); P < 0.009). Ruminal pH did not differ (P = 0.13) and averaged 5.6 +/- 0.06. No clinical cases of PEM were observed during the course of either study. The use of thiamine as a dietary additive to aid in the prevention of PEM in finishing lambs does not appear to be necessary under the conditions of this study.

Influence of pressed beet pulp and concentrated separator by-product on intake, gain, efficiency of gain, and carcass composition of growing and finishing beef steers.

The objectives of this experiment were to determine a NE value for pressed beet pulp and the value of concentrated separator by-product (de-sugared molasses) as a ruminal N source in growing and finishing diets for beef cattle. One hundred forty-four cross-bred beef steers (282 +/- 23 kg of initial BW) were used in 2 experiments (growing and finishing). A randomized complete block design was used, with a 3 x 2 factorial arrangement of treatments (level of pressed beet pulp and inclusion of concentrated separator by-product) for both studies. Steers were blocked by BW and allotted randomly to 1 of 6 treatments. In the growing study, the control diet contained 49.5% corn, 31.5% corn silage, 10.0% alfalfa hay, and 9.0% supplement (DM basis). Pressed beet pulp replaced corn at 0, 20, or 40% of dietary DM, and concentrated separator by-product replaced corn and urea at 10% of dietary DM. The growing study lasted for 84 d. Initial BW was an average of 2-d BW after a 3-d, restricted (1.75% of BW) feeding of 50% alfalfa hay and 50% corn silage (DM basis), and final BW was an average of 2-d BW after a 3-d, restricted (1.75% of BW) feeding of 31.5% corn silage, 10.0% alfalfa hay, 25.0% dry-rolled corn, 20.0% pressed beet pulp, 5.0% concentrated separator by-product, and 8.5% supplement (DM basis). After the growing study, the steers were weighed (415 +/- 32 kg), rerandomized, and allotted to 1 of 6 finishing diets. The control diet for the finishing study included 45% dry-rolled corn, 40% high-moisture corn, 5% brome hay, 5% pressed beet pulp, and 5% supplement. Pressed beet pulp replaced high-moisture corn at 5.0, 12.5, and 20.0% of the dietary DM, and concentrated separator by-product replaced high-moisture corn and supplement at 10.0% of diet DM. Steers were slaughtered on d 83 or 98 of the study. In the growing study, the addition of pressed beet pulp to growing diets linearly decreased (P = 0.001) DMI and ADG and inclusion of 10% concentrated separator by-product decreased (P = 0.001) G:F. Increased levels of pressed beet pulp in the finishing diets caused a linear decrease (P = 0.001) in ADG and tended (P = 0.06 and 0.07 for kg/d and % of BW, respectively) to quadratically decrease DMI, whereas addition of concentrated separator by-product increased (P = 0.02 and 0.001 for kg/d and % of BW, respectively) DMI. Apparent NEg of pressed beet pulp was 94.2% of that of corn in the growing study and 81.5% of that of corn in the finishing study.

Influence of steam-peeled potato-processing waste inclusion level in beef finishing diets: effects on digestion, feedlot performance, and meat quality.

Inclusion of potato-processing waste (PW) from the frozen potato products industry in high-grain beef cattle finishing diets was evaluated in two studies. In a randomized complete block design, 125 crossbred yearling heifers (365 +/- 0.3 kg initial BW; five pens per treatment; five heifers per pen) were used to evaluate PW level on feedlot performance and meat quality. Heifers were fed for 85 (two blocks) or 104 d (three blocks). In a digestion study, four ruminally, duodenally, and ileally cannulated Holstein steers (474.7 +/- 26.6 kg initial BW) were used in a 4 x 4 Latin square design to evaluate effects of PW level on ruminal fermentation, site of digestion, and microbial protein synthesis. The control diet for both studies contained 80% corn, 10% alfalfa hay, 5% concentrated separator by-product (CSB), and 5% supplement (DM basis). Potato waste replaced corn and separator by-product (DM basis) in the diet at 0, 10, 20, 30, and 40% in the feedlot study, and at 0, 13, 27, and 40% in the digestion study. In the feedlot study, DMI decreased (linear; P = 0.007) with increasing inclusion of PW. Increasing PW decreased ADG and feed efficiency from 0 to 30% and then increased at 40% (quadratic; P < 0.01). Calculated dietary NEg concentrations did not differ among treatments (P = 0.18). Hot carcass weight decreased as PW increased from 0 to 30% and then increased at 40% PW (cubic; P < 0.01). Fat thickness and longissimus muscle area decreased with increasing PW (linear; P < 0.05). Level of PW did not affect marbling or liver scores (P > 0.30). No difference (P > 0.20) was observed for Warner-Bratzler shear force at 0, 10, 20, and 30% PW levels; however, 40% PW resulted in lower (P = 0.05) shear force values. Taste panel scores for juiciness and flavor intensity did not differ with increasing PW (P > 0.30). Steaks from cattle fed 0% were scored less tender than 10 and 40% PW (cubic; P < 0.05). In the digestion study, DMI decreased (quadratic; P < 0.01) with increasing PW. Ruminal pH and total VFA concentration increased (linear; P < 0.05) and true N disappearance from the stomach complex and apparent total-tract N disappearance decreased with increasing level of PW (linear; P < 0.01). Starch intake and ruminal disappearance decreased with increasing level of PW (quadratic; P < 0.05). Inclusion of PW decreased feedlot performance, with little effect on carcass characteristics or meat quality. Optimal inclusion of PW in finishing diets may depend on the cost of transportation and other dietary ingredients.

Melanin offers protection against induction of cyclobutane pyrimidine dimers and 6-4 photoproducts by UVB in cultured human melanocytes.

The goal of this investigation was to correlate the melanin content in human pigmentary cells with the generation of UVB-induced photoproducts and to examine the relationship between the melanin content and the removal of the photoproducts. Cultured melanocytes from light-skinned individuals synthesized less melanin and produced more cyclobutane pyrimidine dimers and 6-4 photoproducts upon UVB exposure than did melanocytes from black skin. Tyrosine-stimulated melanogenesis provided protection against DNA damage in both cell types. In another set of pigmented cell lines a ratio between eumelanin and pheomelanin was determined. The assessment of association between DNA damage induction and the quantity and quality of melanin revealed that eumelanin concentration correlated better with DNA protection than pheomelanin. Skin type-I and skin type-VI melanocytes, congenital nevus (CN)-derived cells and skin type-II melanocytes from a multiple-melanoma patient were grown in media with low or high L-tyrosine concentration. The cells were irradiated with 200 J/m2 UVB, and the levels of the photoproducts were determined immediately and after 6 and 24 h. Once again the induction of the photoproducts was mitigated by increased melanogenesis, and it was inversely correlated with the skin type. No significant differences were found for the removal of photoproducts in the cultures of skin types I and VI and CN cells. No indications of a delay in the removal of photoproducts in the melanocytes from the multiple-melanoma patient were found either.

Comparison of immunoaffinity chromatography enrichment and nuclease P1 procedures for 32P-postlabelling analysis of PAH-DNA adducts.

32P-postlabelling analysis for detecting DNA adducts formed by polycyclic aromatic compounds is one of the most widely used techniques for assessing genotoxicity associated with these compounds. In cases where the formation of adducts is extremely low, a crucial step in the analysis is an enrichment procedure for adducts prior to the radiolabelling step. The nuclease P1 enhancement procedure is the most established and frequently used of these methods. An immunoaffinity procedure developed for class specific recognition for polycyclic aromatic hydrocarbon (PAH)-DNA adducts has therefore been compared with the nuclease P1 method for a range of DNA adducts formed by PAHs. The evaluation was carried out with skin DNA from mice treated topically with benzo[a]pyrene, 7,12-dimethylbenz[a]anthracene, 5-methylchrysene or chrysene. The immobilised antibody had the highest affinity for adducts structurally similar to the BPDE-I-deoxyguanosine adduct ([+/-]-N2-(7r,8t,9r-trihydroxy-7,8,9,10-tetrahydrobenzo[a]pyrene-1 0t-yl)-2'-deoxyguanosine) against which the antibody had been raised. Of the PAH-modified DNAs evaluated, the maximum adduct recovery was obtained for DNA containing the BPDE I-deoxyguanosine adduct. With DMBA-modified DNA, the profiles of adducts recovered from the column were similar when the column material was treated either with a digest of DMBA-modified DNA or with 32P-labelled DMBA adducts. I-compounds (endogenous adducts in tissue DNA of unexposed animals), which had similar chromatographic properties to PAH-DNA adducts, were not enriched by the immunoaffinity procedure. Compared to the simple nuclease P1 enhancement procedure, the immunoaffinity methods were lengthier and more labour intensive. Advantages of the immunoaffinity procedure include: specificity, allowing the selective detection of a certain class of adducts: efficient adduct enrichment, providing a viable alternative to other enrichment procedures; adequate sensitivity for model studies and the potential to purify adducts for further characterisation. However, as a general screen for detecting the formation of DNA adducts, the nuclease P1 procedure was viewed as the initial method of choice since it was capable of detecting a wider range of PAH-DNA adducts.

Cell proliferation and DNA adducts in hamster tracheal epithelium exposed to benzo[a]pyrene in organ culture.

The effect of benzo[a]pyrene (B[a]P) on cell proliferation in cultured hamster tracheal epithelium was studied in relation to the formation of B[a]P-DNA adducts. To this end, tracheae were isolated from Syrian golden hamsters, cut into rings and cultured in Ham's F12 medium. Then, the tracheal rings were exposed to 2 or 20 mumB[a]P, either continuously for 7 days or for 2 days followed by a 5-day recovery period without B[a]P. At intervals rings were sampled for determination of cell proliferation (by means of the labelling index). In addition, B[a]P-DNA adduct levels were determined in the continuous exposure experiment by means of in situ detection by immunofluorescence microscopy. After 2 days of exposure to 2 or 20 mum B[a]P, there was a significant increase in B[a]P-DNA adduct level. A further, linear increase in B[a]P-DNA adduct level, however, was only observed after continuous exposure to 20 mum B[a]P, whereas the adduct level in the 2 mum continuous exposure group remained virtually the same. In unexposed tracheal epithelium an initial peak of cell proliferation was observed. This initial proliferation was significantly lower in the exposed samples. Only continuous exposure to 20 mum B[a]P steadily decreased the labelling index from day 2 to 7. It is concluded that the increase in B[a]P-DNA adduct level is correlated with the reduction of cell proliferation in hamster tracheal epithelium exposed to B[a]P in Ham's F12 medium.

Molecular dosimetry of DNA damage induced by polycyclic aromatic hydrocarbons; relevance for exposure monitoring and risk assessment.

Polycyclic aromatic hydrocarbons (PAH) form a large group of organic chemicals that are widely distributed in our environment as pollutants of air, water and soil. Several PAH are carcinogenic in rodents, while exposure to these compounds has been associated with various types of human cancer. Upon entering the body, PAH may be converted into reactive electrophilic species, which can give rise to the formation of DNA adducts. DNA adduct formation is considered to be the initial event in chemical carcinogenesis. In this paper, two methods are illustrated that are widely used to determine PAH-DNA adduct formation, namely 32P-postlabelling, and immunochemical analysis with specific antibodies. The applications of the 32P-postlabelling assay comprise the following: A study of interspecies differences in PAH bioactivation in vitro, with microsomal preparations isolated from liver tissue of various rodent species and of human origin; the results indicate that there are considerable qualitative differences between the adduct patterns obtained, which is relevant with respect to extrapolation from animal to man. The analysis of DNA adduct formation in fish retrieved from marine environments polluted to various extents with PAH; results of these studies show a correlation between liver-DNA adduct levels in these fish and the degree of PAH contamination in the aquatic environment. Biomonitoring of PAH exposure through analysis of adducts in blood cells obtained from heavy and light smokers; the data show a fair correlation between PAH-DNA adduct levels in white blood cells and cotinine content in blood plasma, the latter being used as a marker for exposure to cigarette smoke.(ABSTRACT TRUNCATED AT 250 WORDS)

Detection of DNA adducts in basal and non-basal cells of the hamster trachea exposed to benzo(a)pyrene in organ culture.

DNA adducts were quantified in hamster tracheas exposed to benzo(a)pyrene (BP) in organ culture, in basal as well as in non-basal cells, by in situ detection with an adduct-specific rabbit antiserum (W2/01) and with a mouse monoclonal antibody against human cytokeratins 5 and 8 (RCK102) to identify hamster trachea basal cells. Recognition by W2/01 of the adduct of (+)-anti-7,8-dihydroxy-9,10-epoxide of BP (BP-diolepoxide; BPDE) to deoxyguanosine (dG) was checked on human white blood cells (WBCs) exposed to BP together with 3-methylcholanthrene (3MC)-induced rat-liver microsomes. By comparison with the adduct levels determined by 32P post-labeling, a lower detection limit of about 1 adduct per 10(6) nucleotides could be deduced. Next, tracheal rings were exposed to BP (40 microM) in organ culture for 2 days, then washed and cultured without BP for another 3 days. At different time points epithelial cells were isolated and cytospin preparations made. Staining of BP DNA adducts combined with that of cytokeratin (both visualized with fluorescence) allowed detection of adducts in both basal and non-basal cells in the same preparation. BP DNA adduct formation in basal and non-basal cells after 2 days of exposure to BP was not different. However, on removal of BP the adducts disappeared significantly faster from basal cells than from non-basal cells. The combination of the two antibodies mentioned above thus allows selective determination of BP DNA adduct levels in different cell types. This could be of importance with regard to the involvement of specific cell types in the process of tumor initiation.

Class-specific immunoadsorption purification for polycyclic aromatic hydrocarbon-DNA adducts.

An immunoenrichment procedure has been developed for applications in the detection and identification of a broad range of polycyclic aromatic hydrocarbon (PAH)-DNA adducts at very low abundance. The procedures are based on a monoclonal antibody raised to r-7,trans-8,trans-9-trihydroxy-cis-10-(N2-deoxyguanosyl-5'-phospha te)- 7,8,9,10-tetrahydrobenzo[a]pyrene (anti-BPDE-N2-dG) which has been tested for cross-reactivity towards DNA and proteins (bovine serum albumin, chicken gamma globulin and human globin) covalently modified with a range of PAH diol-epoxides. The antibody recognised DNA adducted with the diol-epoxides of benzo[a]pyrene, benz[a]anthracene, chrysene, dibenz[a,h]anthracene or picene. The antibody also cross-reacted with the 7,8,9,10-tetraol derived from benzo[a]pyrene and the 1,2,3,4-tetraols of benz[a]anthracene and chrysene. The degree of cross-reactivity was greatest for PAH adducts with structural features similar to anti-BPDE-N2-dG proximate to the base attachment. The antibody also recognised a range of PAHs adducted to human globin; these included adducts of benzo[a]pyrene, benz[a]anthracene and chrysene diol-epoxides. This wide range of recognition provides good evidence for the class-specific recognition of PAH adducts by the antibody. When immobilized on Sepharose 4B and used in the immunoadsorption purification of adducted nucleotides, the antibody selectively enriched adducts of benzo[a]pyrene, benz[a]anthracene and chrysene from normal nucleotides. Quantitative measurements with [14C]benzo[a]pyrene-DNA adducts showed that the immobilized antibody was able to enrich benzo[a]pyrene adducts from a DNA hydrolysate containing adducts at a level of 1 adduct/10(10) normal nucleotides. In addition, this immunoadsorption technique was effective in enriching a mixture of DNA adducts formed in the skin of CF1 mice treated cutaneously with a mixture of [3H]benzo[a]pyrene and [3H]chrysene. Class-specific immunoenrichment procedures for DNA adducts are important in assisting the identification of genotoxic components in complex mixtures. The performance characteristics of this immobilized antibody suggest that it may be suitable for application in the detection, identification and monitoring of human exposures to low levels of PAHs.

DNA adducts in hamster and rat tracheas exposed to benzo(a)pyrene in vitro.

Syrian golden hamsters are much more susceptible than Wistar rats to the induction of tracheal tumors by benzo(a)pyrene (BP). In order to investigate whether this difference is reflected in the pattern of DNA-adduct induction and removal, tracheas from either species were isolated and exposed to BP (5 micrograms/ml) in organ culture. At various time-points BP-DNA adducts in the epithelial cells were quantified by 32P-postlabeling; unscheduled DNA synthesis (UDS) was determined by [3H]thymidine incorporation. In an induction-repair experiment tracheas were exposed to BP for 2 days, and cultured for another 4 days without BP. After 2 days of exposure total BP-DNA adduct levels were 10 times higher in hamster compared to rat tracheas. In hamster tracheas one major adduct was formed (95%), vs. the adduct between (+)-anti-BP-diolepoxide and deoxyguanosine (BPDE-N2dG). In rat tracheas BPDE-N2dG comprised about 60% of the total adduct level. During exposure to BP the adduct level in hamster trachea increased to 36 +/- 19 adducts/10(6) nucleotides (add/10(6) n) on day 2. Two days after removal of BP the BP-DNA adduct level had decreased to 60% of that on day 2; there was no further decrease in the BP-DNA adduct level. UDS increased during exposure to BP and decreased after removal of BP. In rats, removal of BP did not lead to a decrease in the BP-DNA adduct level, which agreed with the observed absence of UDS. In a second experiment tracheas were exposed to BP continuously for 15 days. In hamster tracheas the total BP-DNA adduct level increased from 11 +/- 0.7 add/10(6) n after 1 day of exposure to 105 +/- 2 add/10(6) n after 15 days; also UDS increased with increasing exposure until day 11. In rat tracheas no progressive increase in the BP-DNA adduct level was seen. It was concluded that the difference in trachea tumor susceptibility between hamsters and rats exposed to BP correlates with the difference between the 2 species in BP-DNA adduct kinetics in the trachea epithelial cells.

Crambe meal is equivalent to soybean meal for backgrounding and finishing beef steers.

Growth, feed efficiency, and carcass characteristics of 70 crossbred steers fed one of four diets were compared. The four diets differed in source of protein supplementation: 100% soybean meal (SB), 67% SB and 33% crambe meal (CM), 33% SB and 67% CM, and 100% CM. All supplements were fed in isonitrogenous amounts. Steers were fed backgrounding diets (12.9% CP) for 84 d and finishing diets (11.2% CP) for 96 d. Average initial weight was 303.4 kg. Backgrounding gains ranged from 1.38 to 1.41 kg/d (P = .92). Finishing gains ranged from 1.43 to 1.47 kg/d (P = .86). Range in entire-experiment gains was 1.41 to 1.46 kg/d (P = .85). Feed efficiencies were equal; entire-experiment efficiencies averaged .144 (P = .96). Growth and efficiency patterns were the same for all four treatments. No overall treatment differences were detected for the seven carcass variables (P = .26 to .96). Average fat depth, longissimus muscle area, yield grade, and dressing percentage were .95 cm, 84.0 cm2, 2.45, and 61%, respectively. At the protein percentage levels of these diets, CM substituted equally for SB for growth rate, feed efficiency, and carcass characteristics.

Detection of benzo[a]pyrene-DNA adducts in cultured cells treated with benzo[a]pyrene diol-epoxide by quantitative immunofluorescence microscopy and 32P-postlabelling; immunofluorescence analysis of benzo[a]pyrene-DNA adducts in bronchial cells from smoking individuals.

Monoclonal antibodies were raised against the reaction product of benzo[a]pyrene diol-epoxide (BPDE) and deoxyguanosine-5'-monophosphate. The antibodies were used for detection of DNA adducts in situ in BPDE-treated cultured human fibroblasts by immunofluorescence microscopy. Analogue-digital conversion of the fluorescence signal and further image processing allowed measurement of the immunospecific fluorescence in the nuclei of these cells. The results are compared with the adduct levels measured in isolated DNA by 32P-postlabelling. Preliminary results are shown of the application of the immunofluorescence method to the analysis of DNA adducts in bronchial cells obtained from smoking individuals.