Fatty acid profile characterization in colostrum, transition milk, and mature milk of primi- and multiparous cows during the first week of lactation.

The specific fatty acid (FA) profile of colostrum may indicate a biological requirement for neonatal calves. The objective of this study was to characterize the FA profile and yields in colostrum, transition milk, and mature milk in primiparous (PP) and multiparous (MP) cows. Colostrum was milked from 10 PP and 10 MP Holstein cows fed the same pre- and postpartum rations. Milkings (M) 2 to 5 and 12 were respectively termed transition and mature milk. Overall, short-chain FA (C4:0 and C6:0) were 61 and 50% lower in colostrum than mature milk, respectively. A parity by milking interaction was also present, with higher C4:0 for PP cows at M2 and for MP cows at M12. Additionally, higher concentrations of C6:0 were present for PP cows at M2 through M4 and for MP cows at M12. Palmitic (C16:0) and myristic (C14:0) acids were 16% and 27% higher in colostrum than mature milk, respectively. However, total saturated FA remained relatively stable. Branched-chain FA were 13% lower in colostrum than mature milk and higher in PP than MP cows throughout the milking period. The proportion of trans-monounsaturated FA (MUFA) was 42% higher in PP cows throughout the milking period, as well as 15% lower in colostrum than mature milk. In contrast, cis-MUFA and total MUFA were not affected by milking nor parity. Linoleic acid (LA) was 13% higher in colostrum than transition and mature milks, but α-linolenic acid (ALA) did not differ. Consequently, the ratio of LA to ALA was 23% higher in colostrum than mature milk and 25% higher in MP cows. Linoleic acid was also 13% higher in MP cows, whereas ALA was 15% higher in PP cows. Conjugated linoleic acid (CLA, cis-9,trans-11) was 63% higher in PP cows, and no differences between colostrum and mature milk were detected. Overall, polyunsaturated FA (PUFA) from the n-6 and n-3 series were over 25% higher in colostrum compared with transition and mature milk. Milking by parity interactions were present for arachidonic acid (ARA), eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), docosahexaenoic acid (DHA), and total n-3 PUFA, translating to higher proportions in PP cows in M1 to M3, whereas proportions remained relatively stable throughout the milking period in MP cows. Despite increasing milk yields throughout the subsequent milkings, higher yields of EPA, ARA, DPA, and DHA were present in colostrum than in mature milk. Greater proportions and yields of n-3 and n-6 FA in colostrum may translate to specific requirements for newborn calves. Differences were also observed between PP and MP cows and may reflect different nutrient requirements and partitioning.

Fatty acid profile characterization in colostrum, transition milk, and mature milk of primi- and multiparous cows during the first week of lactation.

The specific fatty acid (FA) profile of colostrum may indicate a biological requirement for neonatal calves. The objective of this study was to characterize the FA profile and yields in colostrum, transition milk, and mature milk in primiparous (PP) and multiparous (MP) cows. Colostrum was milked from 10 PP and 10 MP Holstein cows fed the same pre- and postpartum rations. Milkings (M) 2 to 5 and 12 were respectively termed transition and mature milk. Overall, short-chain FA (C4:0 and C6:0) were 61 and 50% lower in colostrum than mature milk, respectively. A parity by milking interaction was also present, with higher C4:0 for PP cows at M2 and for MP cows at M12. Additionally, higher concentrations of C6:0 were present for PP cows at M2 through M4 and for MP cows at M12. Palmitic (C16:0) and myristic (C14:0) acids were 38% and 19% higher in colostrum than mature milk, respectively. However, total saturated FA remained relatively stable. Branched-chain FA were 13% lower in colostrum than mature milk and higher in PP than MP cows throughout the milking period. The proportion of trans-monounsaturated FA (MUFA) was 72% higher in PP cows throughout the milking period, as well as 13% lower in colostrum than mature milk. In contrast, cis-MUFA and total MUFA were not affected by milking nor parity. Linoleic acid (LA) was 25% higher in colostrum than transition and mature milks, but α-linolenic acid (ALA) did not differ. Consequently, the ratio of LA to ALA was 29% higher in colostrum than mature milk and 33% higher in MP cows. Linoleic acid was also 15% higher in MP cows, whereas ALA was 15% higher in PP cows. Conjugated linoleic acid (CLA, cis-9,trans-11) was 2.7-fold higher in PP cows, and no differences between colostrum and mature milk were detected. Overall, polyunsaturated FA (PUFA) from the n-6 and n-3 series were over 40% higher in colostrum compared with transition and mature milk. Milking by parity interactions were present for arachidonic acid (ARA), eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), docosahexaenoic acid (DHA), and total n-3 PUFA, translating to higher proportions in PP cows in M1 to M3, whereas proportions remained relatively stable throughout the milking period in MP cows. Despite increasing milk yields throughout the subsequent milkings, higher yields of EPA, ARA, DPA, and DHA were present in colostrum than in mature milk. Greater proportions and yields of n-3 and n-6 FA in colostrum may translate to specific requirements for newborn calves. Differences were also observed between PP and MP cows and may reflect different nutrient requirements and partitioning.

Physicochemical properties of breast meat from water-stressed naked-neck and Ovambo chickens.

1. Differences in growth performance, physicochemical properties and fatty acid composition of breast muscle from water-stressed 16-week-old naked-neck (NNK) and Ovambo (OVB) chickens were investigated. 2. OVB chickens had superior (P < 0.05) slaughter weights at 16 weeks of age, average daily gain (ADG) and average daily water intake (ADWI) than NNK chickens. Body weights of birds at 16 weeks of age, ADG, average daily feed intake (ADFI), ADWI and water to feed ratio (WFR) declined progressively with increasing severity of water restriction while the opposite was observed for feed conversion ratio (FCR) values. NNK chickens had better FCR at the 40% of ad libitum water intake than OVB chickens. 3. Meat from NNK chickens had higher redness (a*) values at the 40% of ad libitum water intake but lower lightness (L*) and yellowness (b*) values at 70% of ad libitum and ad libitum water intakes compared with OVB chickens. 4. There was no interaction between strain and water intake on most fatty acids, except for the proportion of elaidic acid (C18:1t9) higher in meat from NNK compared with OVB chickens given water at 40% of ad libitum. Water restriction to 40% of ad libitum water intake had a positive influence on the proportions of linoleic acid (C18:2n-6), arachidonic acid (C20:4n-6), adrenic acid (C22:4n-6), docosahexaenoic acid (C22:6n-3), total polyunsaturated fatty acid (PUFA), total omega-3 PUFA and total omega-6 PUFA proportions, but resulted in lower proportions of total monounsaturated fatty acids (MUFAs) compared with 70% of ad libitum and ad libitum water intakes. 5. It was concluded that cooking loss, meat redness values, omega-3 and omega-6 PUFA proportions and n-6/n-3 ratio of NNK chickens improved with increasing severity of water restriction compared with OVB chickens.

Association analyses of single nucleotide polymorphisms in bovine stearoyl-CoA desaturase and fatty acid synthase genes with fatty acid composition in commercial cross-bred beef steers.

Two previously reported non-synonymous coding single nucleotide polymorphisms (SNPs) of bovine stearoyl-CoA desaturase (delta-9-desaturase) (SCD) (c.878C>T) and fatty acid synthase (FASN) (g:17924A>G) were assessed for their associations with 72 individual and 12 groups of fatty acids in brisket adipose tissue of 223 Canadian commercial cross-bred beef steers. It was found that the 'CC' genotype of the SCD SNP was significantly associated with lower concentrations of saturated fatty acids (SFA) including 10:0, 14:0 and 20:0, higher concentrations of monounsaturated fatty acids including 9c-14:1, 12c-16:1 and 13c-18:1, higher concentrations of polyunsaturated fatty acids (PUFA) including 9c,15c-18:2, 10c,12c-18:2, 11c,13t-18:2 and 12c,14t-18:2, but lower concentrations of other PUFA of 9c,13t/8t,12c and 20:2n-6 (P < 0.05). The 'AA' genotype of the FASN SNP was significantly associated with higher concentrations of SFAs of 10:0, 12:0, 13:0, 14:0 and 15:0, lower concentrations of unsaturated fatty acids of 9c-18:1 and 20:3n-6, and higher concentrations of unsaturated fatty acids of 9c-14:1 and 12c-16:1 (P < 0.05). Significant epistatic effects between the SCD and FASN SNP genotypes were also found for several fatty acids including 10:0, 23:0, 6t/7t/8t-18:1, 12t-18:1, 13t/14t-18:1, 16t-18:1, total trans18:1 and 9c,13t/8t,12c-18:2 (P < 0.05). These results further suggest that SCD and FASN are strong candidate genes influencing fatty acid composition in beef cattle.

Increasing omega-3 levels through dietary co-extruded flaxseed supplementation negatively affects pork palatability.

To clarify the impact of feeding co-extruded flaxseed on carcass quality and pork palatability, 96 pigs (48 barrows and 48 gilts) were fed three different levels of flaxseed (0%, 5% and 10% of dietary intake) for 76days. Carcass quality and meat quality characteristics of pure loin muscle and ground pork (20% fat) were evaluated. Fat hardness and belly firmness decreased (P<0.001) with increasing co-extruded flaxseed. Pigs fed co-extruded flaxseed levels had higher lean yield (P=0.045) and total lean content (P=0.034). Loin from barrows had higher fat content compared to gilts (P<0.001). Co-extruded flaxseed supplementation increased (P<0.001) omega-3 content in loin and ground pork. Pork flavour intensity and off-flavour intensity scores lowered (P<0.001) with increasing levels of co-extruded flaxseed, being more accentuated (P=0.023) in reheated pork chops from barrows. Diet affected all texture and flavour sensory characteristics (P<0.05) as tissue levels of omega-3 fatty acids increased, likely as a result of increased lipid oxidation.

Authentication of barley-finished beef using visible and near infrared spectroscopy (Vis-NIRS) and different discrimination approaches.

This study evaluated visible and near-infrared spectroscopy (Vis-NIRS) to authenticate barley-finished beef using different discrimination approaches. Dietary grain source (barley, corn, or blend-50% barley/50% corn) did not affect (P > 0.05) meat quality but influenced (P < 0.05) fatty acid profiles. The longissimus thoracis (LT) from barley-fed steers had lower n-6 fatty acid content and n-6/n-3 ratio compared to LT from corn and blended grain-fed steers. Vis-NIRS coupled with partial least square discriminant analysis (PLS-DA) and support vector machine in the linear (L-SVM) kernel classified with approximately 70% overall accuracy subcutaneous fat and intact LT samples, respectively, from barley, corn, and blended-fed cattle. When only barley and corn samples were considered, fat and intact LT samples were correctly classified with overall accuracy >94% with PLS-DA and radial/L-SVM, and approximately 90% with PLS-DA and L-SVM, respectively. Ground LT samples were classified with ≤70% overall accuracy. Vis-NIRS measurements on fat and intact LT have potential to discriminate between corn and barley-fed beef.

Scanning thermal microscopy and Raman analysis of bulk fused silica exposed to low-energy femtosecond laser pulses.

Low energy femtosecond laser pulses locally increase the refractive index and the hydro-fluoric acid etching rate of fused silica. These phenomena form the basis of a direct-write method to fabricate integrated glass devices that are of particular interest for optofluidics and optomechanical applications. Yet the underlying physical mechanism behind these effects remains elusive, especially the role of the laser polarization. Using Scanning Thermal Microscope and Raman spectrometer we observe in laser affected zones, a localized sharp decrease of the thermal conductivity correlated with an increased presence of low-number SiO(2) cycles. In addition, we find that a high correlation exists between the amount of structural changes and the decrease of thermal conductivity. Furthermore, sub-wavelength periodic patterns are detected for high peak power exposures. Finally, our findings indicate that, to date, the localized densification induced by femtosecond laser pulses remains well below the theoretical value achievable in mechanically densified silica.

UGT1A1 promoter polymorphism increases risk of nilotinib-induced hyperbilirubinemia.

Nilotinib is a novel BCR-ABL inhibitor with significantly improved potency and selectivity over imatinib. In Phase I and Phase II clinical studies of nilotinib in patients with a variety of leukemias, infrequent instances of reversible, benign elevation of bilirubin were observed. Uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1) glucuronidates bilirubin in humans, and a polymorphism in the promoter of the gene that encodes it has been associated with hyperbilirubinemia during treatment with a number of drugs. Pharmacogenetic analysis of that TA-repeat polymorphism found an association between the (TA)7/(TA)7 genotype and risk of hyperbilirubinemia in Phase I patients with imatinib-resistant/intolerant chronic myeloid leukemia (CML) or relapsed/refractory Ph+ acute lymphoblastic leukemia (ALL); this result was replicated in two separate analyses of the chronic phase (CP) and accelerated phase (AP) CML arms of a Phase II study. As nilotinib is not known to be glucuronidated by UGT1A1, the combined impact of inhibition of UGT1A1 activity by nilotinib and genetic polymorphism is the most likely cause of the increased rate of hyperbilirubinemia.

Characterisation of intramuscular, intermuscular and subcutaneous adipose tissues in yearling bulls of different genetic groups.

Fatty acid (FA) composition of intramuscular (IM, Longissimus thoracis muscle), intermuscular (IT) and subcutaneous (SC) fat of one hundred intensively fed yearling bulls with different propensities to fatten were studied. Meat samples were collected from Asturiana de los Valles bulls with different genotypes with respect to the myostatin gene (mh/mh n=24, mh/+ n=26 and +/+ n=25) and from Asturiana de la Montaña (n=25) bulls lacking the mutation responsible for double muscling and characterised by small to medium-frame size adapted to less favoured mountain areas. FA profiles were expressed as percentages of total FA (g/100g of total FA) and organised into groups (saturated (SFA), branched (BFA), monounsaturated (MUFA), C18:1trans, polyunsaturated (PUFA), n-6, n-3, conjugated linoleic acids (CLA), unsaturated (UFA)) and ratios (MUFA/SFA (M/S), PUFA/SFA (P/S), UFA/SFA (U/S), n-6/n-3). The IT depot was the most saturated and SC depot contained the most monounsaturated FAs, while IM fat had the most polyunsaturated FAs. IM fat showed the highest P/S ratio and for the n-6/n-3 ratio there were no significant differences between adipose tissue depots. In general, genotype effects were more pronounced in IM and SC fat profiles compared to the IT depot, for which no significant differences between genotypes were found in SFA, PUFA (including n-6 and n-3), UFA and most of the ratios. IM fat of mh/mh animals had the highest content of PUFA and thus the highest P/S ratio. Accordingly, the presence of the gene causing double muscling influenced the tendency to deposit carcass fat and its FA composition, mainly in IM fat. In general, when carcass fat decreased, SFA content decreased while PUFA and UFA contents increased due to the changes in their percentages.

Phenotypic and genetic correlations of fatty acid composition in subcutaneous adipose tissue with carcass merit and meat tenderness traits in Canadian beef cattle.

Bivariate animal models were used to estimate phenotypic and genetic correlations between 9 carcass merit and meat tenderness traits with 25 individual and grouped fatty acids in the subcutaneous adipose tissue of a population of 1,366 Canadian beef cattle finishing heifers and steers. In general, phenotypic correlations were low (<0.25 in magnitude) except for moderate phenotypic correlations of 9-17:1 (-0.29 ± 0.16), 18:0 (0.26 ± 0.14), 11-18:1 (-0.33 ± 0.15), 11-18:1 (0.35 ± 0.14) with Warner-Bratzler shear force measured 3 d postmortem and between 14:0 (-0.36 ± 0.1), 9-14:1 (-0.34 ± 0.08), 9-16:1 (-0.36 ± 0.08), 9-18:1 (0.26 ± 0.07), and sum of branched-chain fatty acids (BCFA; -0.27 ± 0.06) and back fat thickness (BFAT). Genetic correlations were also low for most of the traits. However, moderate to moderately high genetic correlations (0.25 to 0.50 in magnitude) were detected for some traits, including 17:0 (0.4 ± 0.11), 18:0 (0.44 ± 0.12), 9-14:1 (-0.47 ± 0.11), 9-16:1 (-0.43 ± 0.11), and the -6:-3 PUFA ratio (-0.5 ± 0.15) with HCW; 9-14:1 (-0.41 ± 0.13) and 9-16:1 (-0.42 ± 0.13) with BFAT; 17:0 (0.43 ± 0.19) and BCFA (0.45 ± 0.19) with lean meat yield; 13-18:1 (0.40 ± 0.15) with carcass marbling score; sum of CLA (0.45 ± 0.22), 18:2-6 (0.47 ± 0.17), and sum of PUFA (0.48 ± 0.17) with overall tenderness measured 3 d postmortem; the -6:-3 PUFA ratio (0.41 ± 0.22) and sum of CLA (0.42 ± 0.25) with overall tenderness measured 29 d postmortem; and BCFA (0.41 ± 0.27) with Warner-Bratzler shear force measured 29 d postmortem. The genetic correlations observed in this study suggest that contents of some fatty acids in beef tissue and carcass merit and meat tenderness traits are likely influenced by a subset of the same genes in beef cattle. Due to some antagonistic genetic correlations, multiple-trait economic indexes are recommended when fatty acid composition, carcass merit, and meat tenderness traits are included in the breeding objective.

Influence of cooking methods and storage time on lipid and protein oxidation and heterocyclic aromatic amines production in bacon.

This study aimed to examine the influence of cooking methods and pre-determined refrigerated storage days on the production of lipid oxidation (TBARS), protein oxidation (PROTOX) and heterocyclic aromatic amines (HAA) in bacon. Forty-four pork bellies selected from pigs varying in breed, sex and diets to introduce variability in composition were processed as bacon. Sliced-bacon was stored at 4°C either for 2 or 28days and these storage groups were cooked either with microwave or frying pan. Microwave led to significantly higher PROTOX (P<0.001), while frying pan led to higher levels of HAA and TBARS in bacon (P<0.001). Frying pan cooking increased the saltiness and crispiness of bacon (P<0.05) whereas other sensory attributes were not affected (P>0.05) by the cooking methods and storage times. Similarly, the fatty acid composition of pork belly did not significantly influence the production of HAA, TBARS and PROTOX produced in bacon during cooking. Overall, microwave cooking had lesser impact on the production of carcinogenic compounds in bacon with only minor impact on sensory attributes.

Compositional and dimensional factors influencing pork belly firmness.

We explored various dimensional and compositional factors that can influence the perception of pork belly firmness. Bellies from 198 pigs of three different genotypes, two sexes, two slaughter weights and three different diets were recovered and belly firmness was assessed using the belly-flop angle and a 5-point scale subjective measurement. Dimensional and compositional factors were recorded on intact and sheet-ribbed bellies. Subjective belly score was negatively correlated with belly-flop angle (r=-0.89). Regression analysis accounted for 77 and 83% of the variability in subjective belly firmness and belly-flop angle measurement, respectively. Belly length, weight and width influenced both measures of belly firmness, but these effects were more important for the belly-flop angle. After correcting flop angle using belly length, the effect of belly weight disappeared and the effect of other traits was more like those observed for subjective scoring. Hence, undue effect of belly length should be corrected for if this set-up is to be implemented in commercial plants.

Phenotypic and genetic relationships of residual feed intake measures and their component traits with fatty acid composition in subcutaneous adipose of beef cattle.

Feed efficiency is of particular interest to the beef industry because feed is the largest variable cost in production and fatty acid composition is emerging as an important trait, both economically and socially, due to the potential implications of dietary fatty acids on human health. Quantifying correlations between feed efficiency and fatty acid composition will contribute to construction of optimal multiple-trait selection indexes to maximize beef production profitability. In the present study, we estimated phenotypic and genetic correlations of feed efficiency measures including residual feed intake (RFI), RFI adjusted for final ultrasound backfat thickness (RFIf); their component traits ADG, DMI, and metabolic BW; and final ultrasound backfat thickness measured at the end of feedlot test with 25 major fatty acids in the subcutaneous adipose tissues of 1,366 finishing steers and heifers using bivariate animal models. The phenotypic correlations of RFI and RFIf with the 25 individual and grouped fatty acid traits were generally low (<0.25 in magnitude). However, relatively stronger genetic correlation coefficients of RFI and RFIf with PUFA traits including the -6:-3 ratio (0.52 ± 0.29 and 0.45 ± 0.31, respectively), 18:2-6 (0.45 ± 0.18 and 0.40 ± 0.19, respectively), -6 (0.43 ± 0.18 and 0.38 ± 0.19, respectively), PUFA (0.42 ± 0.18 and 0.36 ± 0.20, respectively), and 9-16:1 (-0.43 ± 0.20 and -0.33 ± 0.22, respectively) were observed. Hence, selection for low-RFI or more efficient beef cattle will improve fatty acid profiles by lowering the content of -6 PUFA, thus reducing the ratio of -6 to -3 along with increasing the amount of 9-16:1. Moderate to moderately high genetic correlations were also observed for DMI with 9-14:1 (-0.32 ± 0.17) and the sum of CLA analyzed (SumCLA; -0.45 ± 0.21), suggesting that selection of beef cattle with lower DMI will lead to an increase amount of 9-14:1 and SumCLA in the subcutaneous adipose tissue. However, unfavorable genetic correlations were detected for ADG with 11-18:1 (-0.38 ± 0.23) and SumCLA (-0.73 ± 0.26), implying that selection of beef cattle with a better growth rate will decrease the contents of healthy fatty acids 11-18:1 and SumCLA. Therefore, it is recommended that a multiple-trait selection index be used when genetic improvements of fatty acid composition, feed efficiency, feed intake, and growth are important in the breeding objective.

Sequential Feeding of Lipid Supplement Enriches Beef Adipose Tissues with 18:3n-3 Biohydrogenation Intermediates.

The present study was designed to determine if feeding steers extruded flaxseed and hay (25 and 75%; DM basis) together as a total mixed ration (TMR), or sequentially (non-TMR) would result in different enrichments of polyunsaturated fatty acids (PUFA) and their biohydrogenation intermediates (BHI) in beef adipose tissues [subcutaneous (SC) vs perirenal (PR) fat]. Forty-eight Angus cross steers (325 ± 16 kg) were stratified by weight to six pens, and pens were randomized to either TMR or non-TMR and fed ad libitum for an average of 242 days. The concentrations of α-linolenic acid increased by 18 mol% in both SC and PR in non-TMR steers compared to TMR steers (P < 0.01). trans 18:1 isomers were more concentrated in PR than SC (14.4 vs 9.5 mol%; P < 0.01) and increased by 10 mol% in both fat depots for non-TMR (P < 0.01). Other BHI including non-methylene-interrupted 18:2 (atypical dienes), conjugated linoleic acids and conjugated linolenic acids (CLnA) were affected by diet × tissue interactions (P < 0.01). The CLnA and CLA contents were higher in both fat depots when feeding the non-TMR, but the effect of diet was more pronounced in PR than in SC (P < 0.01). Atypical dienes were highest in PR from non-TMR and lowest in TMR fed steers (4.3 and 3.6 mol%) with SC contents being intermediate. The sequential feeding of lipid supplement can thus profoundly affect the enrichment of PUFA and their BHI in beef fat and their differentially enrichment is also fat depot dependant.

Effects of feeding steers extruded flaxseed on its own before hay or mixed with hay on animal performance, carcass quality, and meat and hamburger fatty acid composition.

The objective of the present experiment was to determine if carcass quality and fatty acid profiles of longissimus thoracis (LT) and hamburger would be affected by feeding steers extruded flaxseed on its own followed by hay (non-TMR) compared to when hay and extruded flaxseed were fed together (TMR). Forty-eight steers in six pens were assigned to TMR or non-TMR for an average of 242days. Dry matter intake was lower for non-TMR versus TMR steers (10.56 vs. 11.42kg/d; P=0.02), but final live weight (610±0.50kg) and average daily gain (1.18±0.02kg/d) did not differ. Compared to TMR, feeding non-TMR enriched LT and hamburger with α-linolenic acid (ALA; 18:3n-3) by 14%, vaccenic acid (VA; t11-18:1) by 44%, rumenic acid (RA; c9,t11-18:2) by 40%, and conjugated linolenic acid (CLnA) by 58%. Overall, feeding extruded flaxseed separately from hay in a non-TMR was more effective at enhancing deposition of ALA, VA, RA and CLnA in beef.

A mapping method for the description of Warner-Bratzler shear force gradients in beef Longissimus thoracis et lumborum and Semitendinosus.

A novel approach to mapping Warner-Bratzler shear of whole muscles was explored. The procedure was used on the Longissimus thoracis et lumborum (LTL) and Semitendinosus (ST) from six beef cattle by first marking skeletally defined anatomical landmarks on the muscle in situ. After removal from the carcass, further divisions were made while preserving sample orientation during cooking and preparation for shearing. Shear gradients were observed in all planes of the LTL, particularly the medial-lateral. The mid-section of the ST had the lowest shears while superficial locations of the cross-section had greater values. Muscle comparison indicated the ST was more uniform than the LTL. The mapping technique was subsequently used to identify localized effects of altered carcass suspension on shear values and sarcomere length in the lumbar Longissimus from four beef cattle. This mapping method will provide guidance for further intensive investigation across the carcass musculature and under varying carcass conditions.

Induction of apoptosis by arachidonic acid in chronic myeloid leukemia cells.

The hallmark of chronic myeloid leukemia (CML) is the presence of the bcr-abl oncogene, which is associated with transforming ability and an intrinsic resistance to induction of apoptosis by genotoxic agents. Arachidonic acid (AA), a biologically active fatty acid, plays a crucial role as a mediator of signaling pathways involved in cell proliferation and survival. In this study, we investigated the potential role of AA as a proapoptotic agent in CML. Pretreatment of human CML isolated progenitor cells with AA (100 microM for 18 h) induced 71-75% inhibition of in vitro colony formation of granulocyte-macrophage colony-forming units, multilineage colony-forming units, and erythroid burst-forming units. This inhibition was significantly greater than the effect on normal progenitor cells (19-39% growth inhibition of erythroid burst-forming units, multilineage colony-forming units, and granulocyte-macrophage colony-forming units). AA also inhibited growth of the bcr-abl-transformed cell line H7.bcr-abl A54. In contrast, a minimal effect of AA on inhibition of cell growth was observed in the parental nontransformed NSF/N1.H7 cell line. The antiproliferative effect of AA was associated with apoptosis. Gamma-linolenic acid, a precursor of AA, also inhibited cell growth, whereas other unsaturated and saturated fatty acids had no effect. Pharmacological inhibition of cyclooxygenase, lipooxygenase, and cytochrome P450 monooxygenase enzymes prior to exposure to AA did not rescue cells from the inhibitory effect of AA. Moreover, 5,8,11,14-eicosatetraynoic acid, a nonmetabolizable arachidonate analogue, also inhibited cell growth, suggesting that the effect of AA did not require further metabolism. Treatment with antioxidants prior to stimulation with AA was also ineffective in preventing its antiproliferative effect. Thus, AA inhibited proliferation of CML cells by inducing apoptotic cell death. The signaling mechanisms of AA-induced inhibition of cell growth appeared to be independent of its conversion into eicosanoids or free radical generation.

Frequent breakpoints in the 3p14.2 fragile site, FRA3B, in pancreatic tumors.

FRA3B, at chromosomal band 3p14.2, is the most active common fragile site in the human genome. Homozygous deletions in the region of FRA3B have been observed in many types of solid tumors. Recently, the FHIT gene was reported to span FRA3B and was shown to be homozygously deleted in several gastric and colonic tumor cell lines. Several microsatellite markers that precisely define the 1.0-Mb region surrounding FRA3B and FHIT have been utilized, along with other 3p microsatellites, to analyze the loss of 3p sequences in 25 primary pancreatic adenocarcinomas. The high density of microsatellite markers in the 3pl4.2 region enabled us to both identify losses within and flanking FRA3B in pancreatic cancer and define the breakpoints. We observed loss of het erozygosity of 3pl4.2 markers in 16 of 25 pancreatic tumors and loss of heterozygosity of 3p markers outside of 3pl4.2 in only 2 of 25 tumors of this type. There appears to be a dramatic clustering of chromosomal breakpoints at 3pl4.2 in and immediately distal to FRA3B in pancreatic cancer. We detected no homozygous deletions in this region.

Lack of effect of cyclophosphamide on the immunogenicity of a melanoma antigen vaccine.

Melanoma antigen vaccines are a conceptually attractive approach to prevent or delay disease recurrence in patients with surgically resected malignant melanoma. However, the immunogenicity of current vaccines is relatively low. Cyclophosphamide, when given in low doses prior to antigen exposure, is an immunomodulator which has been shown to enhance both humoral and cellular antitumor responses in animals and humans. We conducted a prospective, randomized, clinical trial to study whether pretreatment with cyclophosphamide augments the immunogenicity of a polyvalent, allogeneic, melanoma antigen vaccine in patients with melanoma and low tumor burden. Forty-five patients with resected stage II melanoma (regional metastases) were randomly allocated to treatment with melanoma vaccine or melanoma vaccine plus cyclophosphamide. All patients received the same dose and schedule of vaccine immunizations; those randomized to cyclophosphamide received 300 mg/m2 i.v. 3 days prior to each vaccine immunization. Cellular immune responses were evaluated by delayed-type hypersensitivity (DTH) skin reactivity to a test dose of vaccine at baseline (prior to treatment) and following the fourth immunization. Humoral immune responses were measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiographic analysis of indirect immunoprecipitates of patients' sera at the same time points. Twenty-four patients were randomized to cyclophosphamide pretreatment and 21 to vaccine alone; 22 and 18 patients were evaluable in each group, respectively. Differences were statistically nonsignificant with respect to either cellular (DTH) or humoral (antibody) responses between the two groups. DTH responses were induced in 16 of 22 (73%) and 15 of 18 (83%) patients treated with cyclophosphamide plus vaccine and vaccine alone, respectively. The mean posttreatment augmentation in DTH response in the cyclophosphamide group was 9.5 mm, compared with 9.9 mm in the vaccine-only group. Eight of 12 (66%) cyclophosphamide-pretreated patients and 9 of 12 (75%) vaccine-only patients produced increased titers of antimelanoma antibodies following treatment. No differences were observed between the groups in disease-free or overall survival. In summary, low-dose cyclophosphamide pretreatment failed to augment the immunogenicity of a polyvalent, allogeneic, melanoma vaccine in patients with completely resected early-stage melanoma.

Non-conjugated cis/trans 18:2 in Beef Fat are Mainly Δ-9 Desaturation Products of trans-18:1 Isomers.

Human liver cells (HepG2) were cultured with individual trans (t) 18:1 including t6-, t12-, t13-, t14-, t15- and t16-18:1, and retention times of their Δ-9 desaturation products were determined using 100-m biscyanopropyl-polysiloxane and SLB-IL111 columns. Corresponding peaks were found in beef adipose tissues known to have different delta-9 desaturase activities. Further lines of evidence indicating the presence of Δ-9 desaturation products of t-18:1 isomers in beef fat were developed by analysis of fatty acid methyl esters (FAME) fractionated using Ag+-TLC, and by GC/MS. Some of the Δ-9 desaturation products of t-18:1 have been previously identified in ruminant fat (c9, t12- and c9, t13-18:2). Some of the Δ-9 desaturation products of t-18:1 (c9, t14- and c9, t15-18:2) have been previously tentatively identified as different fatty acids, and for the first time we provide evidence of the presence of c9, t16-18:2, and where t6, c9-18:2 may elute during analysis of FAME from beef fat.