Lokenga Badinga

Recent interest in conjugated linoleic acid (CLA) research stems from the well-documented anticarcinogenic, antiatherogenic, antidiabetic, and antiobesity properties of CLA in animal models. The objective of this study was to examine the effects of 2 CLA isomers (cis-9,trans-11 and trans-10,cis-12) on phorbol 12,13-dibutyrate (PDBu)-induced PGF2alpha production in cultured bovine endometrial (BEND) cells. Confluent BEND cells were incubated in the absence (control) or presence of 100 microM each of linoleic acid, cis-9,trans-11 CLA, or trans-10,cis-12 CLA for 24 h. After incubation, cells were rinsed and then stimulated with PDBu (100 ng/mL) for 6 h. Compared with untreated cells, PDBu stimulated PGF2alpha secretion (+25-fold) within 6 h. The increases in PGF(2alpha) secretion were paralleled by signifi-cant induction of prostaglandin endoperoxide synthase-2 (PGHS-2) mRNA (+63-fold) and protein (+1.6-fold) expression. In spite of stimulatory effects on PGHS-2 and peroxisome proliferator-activated receptor delta (PPARdelta) mRNA responses, CLA greatly decreased PGF2alpha production by PDBu-stimulated BEND cells. There was no evidence for PDBu or CLA modulation of PPARdelta protein synthesis in cultured BEND cells. Results indicated that CLA modulation of PGF2alpha production by BEND cells was not mediated through PGHS-2 or PPARdelta gene repression.

Conjugated linoleic acid (CLA) refers to a mixture of positional and geometric dienoic isomers of linoleic acid found naturally in animal products of ruminant sources. Recent interest in CLA research stems from the well‐documented anticarcinogenic, antiatherogenic, antidiabetic, and antiobesity properties of CLA in rodents. However, there has been very little published human research on CLA. This review discusses the physiologic properties of CLA and their potential implications for human health.

Recent studies have implicated n-3 polyunsaturated fatty acids in the reduction of eicosanoid production in the bovine uterus. The objective of this study was to determine whether the effect of eicosapentaenoic acid (EPA; C(20:5), n-3) on PGF2alpha production by bovine endometrial (BEND) cells is influenced by the quantity of linoleic acid (C(18:2), n-6) in the incubation medium. Confluent BEND cells were incubated in the absence (control) or presence of 100 microM of EPA for 24 h. After incubation, cells were rinsed and then stimulated with phorbol 12,13-dibutyrate (PDBu; 100 ng/mL) for 6 h. Additional sets of culture dishes were treated with a combination of EPA and increasing n-6/n-3 fatty acid ratios for 24 h and then challenged with PDBu for 6 h. The PDBu stimulated PGF2alpha secretion and upregulated steady-state concentrations of prostaglandin endoperoxide synthase-2 and peroxisome proliferator-activated receptor delta mRNA within 6 h. Preincubation of BEND cells with EPA for 24 h decreased PGF2alpha response to phorbol ester, but had no detectable effects on prostaglandin endoperoxide synthase-2 or peroxisome proliferator-activated receptor delta mRNA abundance in PDBu-stimulated BEND cells. The inhibitory effect of EPA on PGF2alpha production was reverted in BEND cells treated with an increasing n-6-to-n-3 fatty acid ratio. Findings indicate that the net inhibition of endometrial PGF2alpha bioynthesis by n-3 fatty acids may vary depending on the ratio of n-6 to n-3 fatty acids in the uterus.

Multiparous Holstein cows, averaging 80 d in milk, were used to examine the effect of exogenous bovine somatotropin (bST) on uterine expression of estrogen receptor alpha (ERalpha), prostaglandin endoperoxide synthase-2 (PGHS-2), and peroxisome proliferator-activated receptor delta (PPARdelta). About 12 h before expected ovulation in a synchronization protocol, cows were assigned to receive bST (500 mg, n = 11) or serve as untreated controls (n = 10). Cows that ovulated (n = 9 bST, 8 control) were divided within treatment to be killed on d 3 or 7 postovulation. Samples of intercaruncular endometrial tissue from uterine horns ipsilateral to the corpus luteum were collected and stored at -80 degrees C for subsequent mRNA analyses. Endometrial concentrations of ERalpha and PGHS-2 mRNA transcripts were greater on d 7 than on d 3 of the estrous cycle, but did not differ between treatments. Compared with untreated cows, short-term bST treatment decreased PGHS-2 protein expression at d 7 of the estrous cycle. Concentration of PPARdelta mRNA transcript in the uterus decreased between d 3 and 7 of the estrous cycle and was negatively correlated with ERalpha and PGHS-2 mRNA concentrations. Short-term administration of bST to lactating dairy cows had minimal effects on uterine genes encoding ERalpha, PGHS-2, and PPARdelta at d 3 and 7 of the estrous cycle but there may be an inverse relationship between PPARdelta and uterine expression of ERalpha and PGHS-2 genes.

Effects of feeding calcium salts of conjugated linoleic acid (CLA) or trans octadecenoic acids (trans 18[ratio ]1) on lipid metabolism and hepatic contents of mRNA encoding carnitine palmitoyltransferase 1 (CPT1), microsomal triglyceride transfer protein (MTP) and peroxisome proliferator-activated receptor alpha (PPARα) were examined in 15 early post-partum Holstein cows. Dietary treatments were initiated at approximately 4 weeks prior to expected calving dates and continued for 7 weeks post partum. Treatments prepartum consisted of 1) a basal diet (Control), 2) basal diet+150 g/d of CLA mix (CLA), or 3) basal diet+150 g/d of trans 18[ratio ]1 mix (TRANS). Intakes of calcium salts of CLA and trans 18[ratio ]1 mixes were adjusted to 225 g/d during the 7-week post-partum treatment period. Blood samples were collected at weeks 1, 2 and 4 post partum and plasma was harvested immediately for subsequent hormone and metabolite assays. Concentrations of insulin, insulin-like growth factor-I (IGF-I), and leptin in blood did not vary among cows fed the three diets. Plasma nonesterified fatty acid (NEFA) concentrations decreased between weeks 1 and 4 of lactation and were lower in cows fed the diet supplemented with trans 18[ratio ]1 than in those fed a control diet at week 2 post partum. Periparturient fat supplementation had no detectable effects on CPT1 mRNA content in the liver. Steady-state concentration of MTP mRNA in the liver was greater in the TRANS treatment group than in the control group at week 1 post partum. Feeding trans 18[ratio ]1 supplements to transition dairy cows upregulated hepatic PPARα mRNA content during the first month of lactation. Under the present experimental conditions, dietary CLA had minimal effects on plasma and hepatic lipid metabolite concentrations in early lactation Holstein cows. Results indicate that dietary trans fatty acids may affect liver lipid metabolism in post-partum dairy cows through alterations in PPARα gene expression.

Thirty-eight multiparous Holstein cows were utilized in a completely randomized design to examine the effect of feeding calcium salts of conjugated linoleic acid (CLA) and trans-octadecenoic acids (trans-C18:1) on animal performance and lipid and glucose metabolism during the transition to lactation. Dietary treatments were initiated approximately 28 d prior to expected calving dates and continued through d 49 postpartum. Prepartum treatments consisted of 1) a basal diet (Control), 2) basal diet + 150 g/d of CLA mix (CLA), and 3) basal diet + 150 g/d of trans-C18:1 mix (TRANS). Amounts of calcium salts of CLA and trans-C18:1 mixes were adjusted to 225 g/d during the 49-d postpartum treatment period. All diets were offered as a total mixed ration. Prepartum fat supplementation had no detectable effects on dry matter intake, body weight, or body condition score. After parturition, cows in the TRANS group consumed less dry matter at wk 4, 5, and 6 of lactation than did cows in the control group. Cows fed the trans-C18:1 supplement were in a more severe negative energy balance than those fed the control diet at 1 wk of lactation. Periparturient fat supplementation had no detectable effects on milk yield during wk 1 to 7 of lactation. Milk fat was not affected during wk 1 to 4, but was reduced after wk 4 of lactation by dietary CLA. Feeding calcium salts of CLA decreased short- to medium-chain fatty acid (C4 to C14) concentrations and increased both linoleic and linolenic acid concentrations in milk fat. Concentrations of nonesterified fatty acids and beta-hydroxybutyric acid in blood were greater in cows fed the CLA-supplemented diet than in those fed the control diet at 1 wk of lactation. In spite of small numerical tendencies, hepatic lipid and triacylglycerol concentrations did not vary significantly among dietary treatments. Periparturient fat supplementation had no detectable effects on plasma glucose and insulin concentrations. Steady-state concentrations of hepatic mRNA encoding pyruvate carboxylase and phosphoenolpyruvate carboxykinase were greater for the TRANS treatment group than the control and CLA groups. Results indicate that dietary CLA and trans-C18:1 fatty acids may affect lipid and glucose metabolism in early postpartum Holstein cows through distinct mechanisms.

We examined the effect of dietary conjugated linoleic acid (CLA) on growth performance and liver composition in broiler chickens. Day-old male broiler chicks were assigned to receive a diet supplemented with corn oil (5%; n = 48) or CLA (5%; n = 48) for 21 d. Broilers fed CLA weighed less and grew at slower rates than broilers fed corn oil. Feed intake and feed conversion were higher for the corn-oil group than for the CLA dietary group. Hepatic lipid and triacylglycerol concentrations were significantly reduced by dietary CLA. The proportions of saturated fatty acids (SFA) in liver lipids increased, whereas those of monounsaturated fatty acids (MUFA) decreased with CLA supplementation. Although the total concentration of polyunsaturated fatty acids (PUFA) did not change with dietary treatment, the concentration of linoleic acid as a percentage of total methylated fatty acids decreased, and that of linolenic add increased in broilers fed CLA. The concentration of CLA isomers in liver lipids increased substantially with CLA feeding. The relative proportion of the c9,t11 CLA isomer in hepatic lipids was much higher than that of the t10,c12 or t9,t11 CLA isomers. These studies provide evidence that feeding CLA to broilers results in substantial reduction in liver fat accumulation and promotes CLA incorporation into hepatic lipid pools.

Lactating Holstein cows, averaging 80 d in milk, were used to examine effects of exogenous bovine somatotropin (bST) on oviductal and uterine genes encoding components of the insulin-like growth factor (IGF) system. About 12 h before expected ovulation in an Ovsynch protocol, cows were assigned randomly to receive bST (500 mg; n = 11) or serve as untreated controls (n = 10). Cows that ovulated (n = 9 bST, 8 control) were divided within treatment to be sacrificed on d 3 or 7 postovulation. Samples of oviductal and intercaruncular endometrial tissue from oviducts and uterine horns ipsilateral to the corpus luteum (CL) were collected and immediately frozen at -80 degrees C for subsequent mRNA analyses. Northern blots revealed mRNAs for IGF-II, IGF-binding protein-2 (IGFBP-2), and IGFBP-3 in all oviductal and endometrial tissues. Significant amounts of IGF-I and growth hormone receptor-1A (GHR-1A) mRNAs were detected in uteri but not in oviducts. The bST treatment had no effect on amount of IGF-I mRNA transcript in uterine endometrium. The mRNA encoding IGF-II was induced by bST in oviducts collected on both d 3 and 7 but was down-regulated in endometrium on d 7. Transcript of IGFBP-2 mRNA was greater in endometrial than oviductal tissues and did not differ between treatments. Both oviductal and endometrial IGFBP-3 mRNA concentrations increased between d 3 and 7 postovulation, with a tendency for greater endometrial IGFBP-3 mRNA in bST-treated cows on d 7. On d 7, concentrations of endometrial GHR-1A mRNA were 30% lower in bST-treated cows. Results indicate complex and tissue-specific regulation of the uterine IGF system components by exogenous bST. Some of those biological responses to bST may be important in early development of bovine embryos.

The recent observation that bovine somatotropin (bST) treatment at a timed insemination improves pregnancy rates in lactating dairy cows raises the possibility that growth hormone (GH) may modulate the endocrine and biochemical cross talk between the conceptus and maternal uterus at the time of pregnancy establishment in cattle. The objective of this study was to characterize the cellular and molecular mechanisms by which exogenous GH affects phorbol ester-induced prostaglandin F2alpha (PGF2alpha) production in cultured bovine endometrial (BEND) cells. Serum-deprived BEND cells were incubated with or without recombinant bovine GH (rbGH), insulin-like growth factor (IGF)-I, recombinant bovine interferon (rbIFN)-tau or a combination of rbGH + rbIFN-tau for 3 h and then treated with phorbol 12,13-dibutyrate (PDBu) for an additional 6 h. Exogenous PDBu increased PGF2alpha secretion and steady-state levels of COX-2 mRNA within 3 h. Priming of BEND cells with rbGH reduced PGF2alpha response to PDBu, whereas cotreatment with IGF-I amplified PDBu induction of PGF2alpha. Preincubation of cell monolayers with rbIFN-tau suppressed PGF2alpha and COX-2 mRNA responses to PDBu. Inhibitory effects of rbGH and rbIFN-tau on PDBu-induced PGF2alpha production were additive. Results provide the first direct evidence that supplemental bST may interact with conceptus-secreted IFN-tau to modulate PGF2alpha secretion at the critical time of maternal recognition of pregnancy.

Eight lactating Holstein dairy cows (80 d in milk) were used to examine the effects of exogenous bovine somatotropin (bST) on hepatic contents of mRNA encoding pyruvate carboxylase (PC), phosphoenolpyruvate carboxykinase (PEPCK), and microsomal triglyceride transfer protein (MTP). Concentrations of bST in plasma were higher and milk production increased 20% in bST-treated cows. Liver samples from cows treated with bST had significantly higher total lipid contents than those from control cows. Although there were small numerical tendencies, neither triglyceride concentrations in liver nor nonesterified fatty acids (NEFA), beta-hydroxybutyrate (BHBA), or glucose in plasma differed significantly between bST-treated and control cows. Short-term bST treatment had no detectable effects on contents of PC, PEPCK, and MTP mRNA in the liver. In summary, exogenous bST stimulation of milk production is not mediated through enhanced liver gluconeogenesis, but may involve partitioning of glucose and fatty acids for preferential use by the mammary gland.