Buffering agent via insulin-mediated activation of PI3K/AKT signaling pathway to regulate lipid metabolism in lactating goats.

Ruminants are often fed a high-concentrate (HC) diet to meet lactating demands, yet long-term concentrate feeding induces subacute ruminal acidosis (SARA) and leads to a decrease in milk fat. Buffering agent could enhance the acid base buffer capacity and has been used to prevent ruminant rumen SARA and improve the content of milk fat. Therefore, we tested whether a buffering agent increases lipid anabolism in the livers of goats and influences of milk fat synthesis. Twelve Saanen-lactating goats were randomly assigned to two groups: one group received a HC diet (Concentrate: Forage=60:40, Control) and the other group received the same diet with a buffering agent added (10 g sodium butyrate, C(4)H(7)NaO(2); 10 g sodium bicarbonate, NaHCO(3); BG) over a 20-week experimental period. Overall, milk fat increase (4.25+/-0.08 vs. 3.24+/-0.10; P<0.05), and lipopolysaccharide levels in the jugular (1.82+/-0.14 vs. 3.76+/-0.33) and rumen fluid (23,340+/-134 vs. 42,550+/-136) decreased in the buffering agent group (P<0.05). Liver consumption and release of nonesterified fatty acid (NEFA) into the bloodstream increased (P<0.05). Phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT) and ribosomal protein S6 kinase (p70S6K) up-regulated significantly in the livers of the buffering agent group (P<0.05). It also up-regulated expression of the transcription factor sterol regulatory element binding protein-1c (SREBP-1c) and its downstream targets involved in fatty acid synthetic, including fatty acid synthetase (FAS), stearoyl-CoA desaturase (SCD-1) and acetyl-CoA carboxylase 1 (ACC1) (P<0.05). The BG diet increased insulin levels in blood (19.43+/-0.18 vs. 13.81+/-0.10, P<0.05), and insulin receptor was likewise elevated in the liver (P<0.05). Cumulatively, the BG diet increased plasma concentrations of NEFA by INS-PI3K/AKTSREBP-1c signaling pathway promoting their synthesis in the liver. The increased NEFA concentration in the blood during BG feeding may explain the up-regulated in the milk fat of lactating goats.

Effect of age on bovine subcutaneous fat proteome: molecular mechanisms of physiological variations during beef cattle growth.

Fat deposition influences both meat quality and animal productivity. However, it is not clear how fat development is regulated in growing and fattening beef cattle. This study characterized proteomic changes in subcutaneous adipose tissue from steers fed a high-grain diet in an effort to understand the molecular mechanisms of fat development during feedlot production. Eight British-Continental crossbred steers had two subcutaneous adipose tissue biopsies at 12 and 15 mo of age. Protein expression in fat samples was profiled using liquid chromatography-tandem mass spectrometry (LC-MS/MS). During the finishing period, steers increased subcutaneous adipose tissue mass with concomitant changes in the proteome profile, but the nature of these changes varied among steers. The expression of 123 out of 627 identified proteins differed (P <: 0.05) between 2 ages. Functional analyses on differentially expressed proteins revealed that 20.2% of them were associated with cellular growth and proliferation of adipose tissue. There were 17 out of 108 differentially expressed proteins associated with lipid metabolism, which were acyl-CoA synthetase medium-chain family member 1 (ACSM1), annexin A1 (ANXA1), apolipoprotein C-III (APOC3), apolipoprotein H (beta-2-glycoprotein I; APOH), EH-domain containing 1 (EHD1), coagulation factor II (thrombin; F2), gelsolin (GSN), lamin A/C (LMNA), mitogen-activated protein kinase kinase 1 (MAP2K1), myosin, heavy chain 9, non-muscle (MYH9), orosomucoid 1 (ORM1), protein disulfide isomerase family A, member 3 (PDIA3), retinol binding protein 4, plasma (RBP4), renin binding protein (RENBP), succinate dehydrogenase complex, subunit A, flavoprotein (Fp; SDHA), serpin peptidase inhibitor, clade C (antithrombin), member 1 (SERPINC1), and serpin peptidase inhibitor, clade G (C1 inhibitor), member 1 (SERPING1). Further analysis of the expression levels of proteins associated with lipid metabolism indicated a downregulation in the synthesis of fatty acids at the cellular level at 15 compared to 12 mo of age. These results suggest that even though adipose tissue expanded, fat anabolism was reduced in adipocytes during growth, revealing a coordinated balance between subcutaneous fat mass and the cellular abundance of lipogenic proteins to control the rate of fat deposition in growing beef cattle. The findings observed in this study expand our understanding on how proteome of bovine adipose tissue is regulated during growth, which might help the development in the future of new strategies to manipulate adiposity in beef cattle in a manner that improves meat quality and animal productivity.

Antitumor effects of interferon-alpha on cell growth and metastasis in human nasopharyngeal carcinoma.

Nasopharyngeal carcinoma (NPC) is a highly malignant and frequently metastasized tumor, and the prognosis is very poor when distant metastases occur. Recently, immunotherapy is becoming a promising therapeutic approach. Interferon-α (IFN-α) represents the cytokines exhibiting the longest record of use in clinical oncology. In this study, we examined the antitumor effects of IFN-α1b on NPC. The results showed that recombinant human IFN-α1b (hIFN-α1b) suppressed cell growth, induced a G1-phase cell cycle arrest in vitro, increased the expression of p16 and pRb, and decreased the expression of CCND1 and CDK6. In vivo analyses showed that either recombinant adeno-associated virus (rAAV)-IFN-α1b or hIFN-α1b treatment inhibited tumor growth and metastasis, reduced intratumoral microvessel density, increased cell apoptosis and necrosis, and induced prolonged survival. Notably, rAAV-IFN-α1b or hIFN-α1b treatment led to significantly higher serum levels of IL-12 and GM-CSF in mice compared to respective controls. Our findings suggest that IFN-α1b acts as a multifunctional antitumor agent in NPC, which may have important therapeutic implications.

Feeding flaxseed in grass hay and barley silage diets to beef cows increases alpha-linolenic acid and its biohydrogenation intermediates in subcutaneous fat.

The objective of the study was to determine temporal fat deposition and fatty acid profiles in beef cows fed hay- or barley silage-based diets, with or without flaxseed. Crossbred cull beef cows (n = 64, >30 mo of age, 620 ± 5 kg) were removed from grassland pastures, randomly assigned to 16 pens, and given ad libitum access to 50:50 (wt/wt, DM basis) forage:concentrate diets containing 0 or 15% ground flaxseed (DM basis, 5.2% added fat). Diets consisted of hay control (HC), hay+flaxseed (HF), barley silage control (SC), and silage+flaxseed (SF). Backfat biopsies were obtained from each cow at 0, 6, and 12 wk, and at slaughter (~20 wk) to assess fatty acid composition. With the exception of feed efficiency, flaxseed × forage interactions were not significant for backfat accumulation or performance parameters. Flaxseed improved (P < 0.01) feed conversion when supplemented to hay-based diet and increased ADG (P = 0.03), resulting in a heavier (P = 0.02) BW. Compared with hay, barley silage increased (P < 0.01) DMI, ADG, and feed efficiency. Subcutaneous fat contained 0.68% n-3 fatty acids at wk 0, and reached 0.68, 0.81, and 0.94% in HF cows after 6, 12, and 20 wk, respectively (Y(n-3) = 0.0133X + 0.6491, r = 0.87). It was 0.67% at wk 0, and reached 0.65, 0.77, and 0.90% in SF cows after 6, 12, and 20 wk, respectively (Y(n-3) = 0.0121X + 0.6349, r = 0.75). In contrast, weight percentage of n-3 fatty acids decreased in HC cows from 0.63, 0.50, and 0.47, to 0.43%, and in SC cows from 0.63, 0.40, and 0.36, to 0.33% over the 20 wk. A forage × flaxseed interaction (P < 0.05) occurred for many of the α-linolenic acid (ALA) biohydrogenation intermediates, including vaccenic acid (C18:1 trans-11) and CLA (combined C18:2 trans-7,cis-9 and cis-9,trans-11) in plasma, and in subcutaneous fat this also included non-CLA dienes. Concentrations of most α-linolenic acid biohydrogenation intermediates were greater when feeding flaxseed with hay. In conclusion, forage source altered plasma concentrations and rate of accumulation of ALA biohydrogenation products in subcutaneous fat from beef cows fed flaxseed. Factors responsible for this response are yet to be defined, but may include forage-mediated changes in ruminal biohydrogenation of ALA, as well as alterations in fatty acid metabolism and deposition.

Cinnamaldehyde in feedlot cattle diets: intake, growth performance, carcass characteristics, and blood metabolites.

Cinnamaldehyde (CIN), a natural chemical compound found in the bark of cinnamon trees, can alter rumen fermentation by inhibiting selected ruminal microbes, and consequently, may improve growth performance and feed efficiency of animals. The objective of this study was to evaluate the effects of supplementing the diet of feedlot cattle with CIN on intake, growth performance, carcass characteristics, and blood metabolites. Seventy yearling steers (BW = 390 +/- 25.2 kg) were assigned to a randomized complete block design with 5 treatments: control (no additive), monensin (MO; 330 mg*steer(-1)*d(-1)), and 400, 800, or 1,600 mg of CIN*steer(-1)*d(-1). At the start of the experiment, steers were blocked according to BW and assigned to 14 blocks of 5 cattle, with cattle within block assigned to treatments. The diets consisted of 9% barley silage, 86% dry-rolled barley grain, and 5% supplement (DM basis). Dry matter intake responded quadratically (P = 0.03) to CIN supplementation with 13% more feed consumed for steers fed CIN (mean of 3 CIN levels) compared with those fed control during the first 28 d of the experiment, and with a tendency of 4% increase over the entire experiment. The ADG (kg/d) tended to respond quadratically (P = 0.08) to CIN supplementation during the first 28 d, but was not affected over the entire experiment (112 d). Feed efficiency (G:F) linearly declined (P = 0.03) during the first 28 d with CIN supplementation and was quadratically affected between d 29 to 56 and d 85 to 112 by CIN dose. Supplementation of MO did not affect (P > 0.15) DMI or growth performance at any time during the experiment. Serum NEFA concentrations were reduced (P = 0.05) by 35, 29, 30, and 22%, respectively, on d 56, 84, 112, and overall with CIN supplementation. Concentrations of serum amyloid A were reduced on d 28 by 56, 60, or 56% for 800 mg of CIN, 1,600 mg of CIN, and MO, respectively, compared with control. Plasma concentrations of lipopolysaccharide binding protein were linearly decreased (P = 0.05) with increasing CIN supplementation on d 28. Results indicate that supplementing a feedlot finishing diet with a small dose of CIN ameliorated feed intake during the initial month but had minimal effects on ADG, feed efficiency, and carcass traits over the entire experiment. Including CIN in the diet of feedlot cattle, particularly early in the feeding period, may help promote intake and reduce the effects of stress.

Carotenoids and prostate cancer risk.

Chemoprevention is presumably one of most effective means to combat prostate cancer (PCa). Patients usually require more than a decade to develop a clinically significant Pca, therefore, an ideal target for chemoprevention. This review will focus on recent findings of a group of naturally occurring chemicals, carotenoids, for potential use in reducing PCa risk.

mRNA expression of gonadotropin-releasing hormones (GnRHs) and GnRH receptor in goldfish.

In goldfish (Carassius auratus), two distinct forms of gonadotropin-releasing hormone (GnRH), namely, salmon GnRH (sGnRH) and chicken GnRH-II (cGnRH-II), have been identified in the brain using chromatographic, immunological, and molecular cloning approaches. These two native GnRHs act on specific receptors in the anterior pituitary to stimulate the synthesis and release of gonadotropins and growth hormone in goldfish. To evaluate the potential roles of sGnRH and cGnRH-II in both neural and reproductive tissues in goldfish, we studied the mRNA expression of sGnRH, cGnRH-II, and GnRH receptor (GnRH-R) in discrete brain areas, pituitary, ovary, and testis by a combined reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analysis. Total RNA was extracted from various tissues of sexually recrudescent male and female goldfish and RT-PCR was performed with primers specific for GnRH-R complementary DNA (cDNA), sGnRH cDNA, cGnRH-II cDNA-1, and cDNA-2. Results showed that GnRHs and GnRH-R mRNAs are differentially distributed in the brain. In the goldfish brain, sGnRH mRNA was predominantly expressed in the forebrain areas (olfactory bulb, telencephalon, and hypothalamus) whereas cGnRH-II mRNA-1 were expressed in all brain areas including olfactory bulbs and optic tectum-thalamus. The expression level of cGnRH-II mRNA-2 was much lower than that of cGnRH-II mRNA-1 in the brain. On the other hand, GnRH-R mRNA was expressed in all brain regions and pituitary. In the ovary and testis, GnRH-R mRNA, sGnRH mRNA, and cGnRH-II mRNA-1, but not cGnRH-II mRNA-2, are expressed. Sequence analysis of the PCR products showed that nucleotide sequences of GnRH-R in gonads are identical with that in the brain and pituitary. The coexistence of GnRHs and GnRH-R mRNAs in both neural and gonadal tissues supports the notion that sGnRH and cGnRH-II may act as neurotransmitters and/or neuromodulators in the brain and as autocrine and/or paracrine hormones in gonadal tissues in addition to their established neuroendocrine roles at the pituitary of goldfish.

Mechanisms of action of growth hormone-releasing peptide-2 in bovine pituitary cells.

We conducted this study to investigate the mechanisms of action of growth hormone-releasing peptide-2 (D-Ala-D-beta Nal-Ala-Trp-D-Phe-Lys-NH2; GHRP-2) in bovine anterior pituitary primary cell culture. Doses of GHRP-2 from 10(-13) to 10(-7) M) increased (P < .05) GH secretion. The GHRP-2 (10(-7) M) and GH-releasing factor (GRF; 10(-7) M) administered together had an additive effect on the release of GH (P < .05). Somatostatin (1 microM) decreased GH secretion in response to GHRP-2 and(or) GRF (P < .05). Secretion of GH in response to GHRP-2 was blocked (P < .01) by a GRF receptor antagonist (.1 microM). Nifedipine (10 microM), a voltage-dependent Ca2+ channel blocker, inhibited (P < .01) GHRP-2-stimulated GH release. The GH release in response to GHRP-2 and 4 beta-phorbol-12-myristate-13-acetate (10(-7) M), a protein kinase C activator, was additive (P < .01). Forskolin (30 microM), a cAMP elevating agent, further stimulated (P < .01) the GH release in response to GHRP-2. Bovine GH concentrations in culture media were assayed by indirect competitive enzyme immunoassay. These results showed that GHRP-2 1) stimulates GH secretion from bovine pituitary cells, 2) may partially act via GRF receptor, 3) has GH secretion activity caused by Ca2+ influx via Ca2+ channels, and 4) may increase GH secretion via protein kinase C and cAMP pathways.

A multicentred phase III comparative clinical trial of Mesigyna, Cyclofem and Injectable No. 1 given monthly by intramuscular injection to Chinese women. I. Contraceptive efficacy and sid effects.

A phase III clinical study was carried out among 5680 fertile Chinese women to evaluate efficacy and side effects of three monthly injectable contraceptives: Mesigyna, Cyclofem and Chinese Injectable No. 1. When used in a once-a-month treatment schedule (part 1 of study), the effectiveness of Chinese Injectable No. 1 was unacceptably low; 36 pregnancies occurred during the first 1743 women-months of use, 16 before the second injection. The study was restarted with a revised injection schedule for Injectable No. 1: two injections separated by 9 +/- 1 days during the first month and subsequent injections given 10-12 days after the onset of bleeding, or if no bleeding occurred, 28 days after previous injection. In part 2 of the study, 988, 990 and 992 subjects were provided Mesigyna, Cyclofem and Injectable No. 1, respectively. Life-table pregnancy rates at one year were 0.41%, 0% and 0.77% (p < 0.05), respectively; the overall discontinuation rates at one year were 13.9%, 19.1% and 20.4% (p < 0.001). Discontinuation rates for bleeding problems were significantly different between the groups: discontinuation rates for amenorrhea were 0.58%, 3.71% and 0.68% (p < 0.001) for Mesigyna, Cyclofem and Injectable No. 1; for other bleeding problems, the rates were 4.88%, 8.38% and 12.64% (p < 0.001). There were no significant differences between the groups regarding discontinuation for other medical or non-medical reasons. Mean weight changes after one year of use were small: 0.73, 0.86 and 0.17 kg for the three groups, respectively. Both Mesigyna and Cyclofem were very effective for contraception, but Mesigyna appeared to be tolerated slightly better with regard to cycle control; the modified dose regimen for Injectable No. 1 also gave a low pregnancy rate but was associated with higher rates of discontinuation.

A multicentred phase III comparative clinical trial of Mesigyna, Cyclofem and Injectable No. 1 given by intramuscular injection to Chinese women. II. The comparison of bleeding patterns.

Between 1988 and 1992, a randomized phase III clinical trial was conducted in China to compare three monthly injectable contraceptives: Mesigyna, Cyclofem and Injectable No. 1. This paper presents a detailed analysis of the menstrual diaries provided by 5098 (89%) of the subjects. In total, 902, 903 and 913 diaries were analyzed to compare bleeding patterns induced by Mesigyna, Cyclofem and Injectable No. 1. The first withdrawal bleeding usually occurs 14-20 days after the first injection for all three of these preparations. Thereafter, 50% of Mesigyna users had precisely 3 bleeding/spotting episodes every 90 days, 50% of Cyclofem users had 2-3 and 50% of Injectable No. 1 users had 3-4 episodes every 90 days. Relative to users of Mesigyna or Cyclofem, Injectable No. 1 users had 2-3 more bleeding/spotting days, and a shorter length of bleeding/spotting-free intervals in each period. 63.7%, 41.4% and 60.6% of subjects using Mesigyna, Cyclofem and Injectable No. 1, respectively, had bleeding patterns similar to their untreated patterns in the first 90-day period. The percentages increased to 82.2% 67.8% and 75.0% in the fourth 90-day period. A total of 1815 diaries for Mesigyna and 1802 for Cyclofem were analyzed for more in depth comparison of these two methods. The number of bleeding/spotting days over four periods showed little difference between the two group, but there were more spotting days and there was greater individual variability among Cyclofem users.(ABSTRACT TRUNCATED AT 250 WORDS)

PIP: Between October 1988 and July 1990, a randomized multicentered phase III clinical trial was conducted in three provinces of China to compare three monthly injectable contraceptives (Mesigyna [50 mg norethisterone enanthate + 5 mg estradiol valerate], Cyclofem [25 mg medroxyprogesterone acetate + 5 mg estradiol cypionate], and Injectable No.1 [250 mg 17-hydroxyprogesterone caproate + 5 mg estradiol valerate]). A detailed analysis of the menstrual diaries of 5098 women aged 18-35 years compared the vaginal bleeding patterns associated with the injectables. Women in all three groups experienced more bleeding/spotting (B/S) days, more bleeding episodes, shorter bleeding-free intervals, and larger variability during the first 90 days than during the following three 90-day periods (p 0.001). 90% of Cyclofem users had 1-4 B/S episodes. 90% of Mesigyna users had 2-4.2 B/S episodes. Cyclofem users had more spotting days than did Mesigyna users in each 90-day period (5-8 vs. 5-6). Acceptable bleeding patterns (i.e., bleeding patterns similar to untreated patterns) predominated, on the most part, in all four periods (63.7-82.2% for Mesigyna users, 41.4-67.8% for Cyclofem users, and 60.6-75% for Injectable No.1 users). Acceptability increased with each 90-day period for all three injectables. Acceptability of bleeding patterns was much higher among Mesigyna users than Cyclofem users (p 0.001). Prolonged bleeding, followed by irregular bleeding and frequent bleeding, were the most common bleeding disturbances. Irregular bleeding decreased with time. 79.1% of Mesigyna and Cyclofem users who finished the study had an acceptable pattern. 70.7% of women who stopped for non-bleeding reasons had an acceptable pattern compared to 31.3% of those who stopped for bleeding reasons. These findings show that Mesigyna users experienced better cycle control and more acceptable bleeding patterns than did the users of the other two injectables.

Effects of different doses of norethisterone on ovarian function, serum sex hormone binding globulin and high density lipoprotein-cholesterol.

The ovarian effects of different doses of norethisterone (NET) were compared in 45 normally menstruating women in order to find the lowest effective dose of the Chinese NET "visiting pill". Subjects were randomly divided into 3 groups. Each subject in each group was taking 0.5, 1.5 or 3.0 mg per day from days 5 to day 18 of the cycle. Blood samples were taken on days 5, 8, 11, 14, 17, 20, 23, 26 and 29 of the cycle. Serum oestradiol (E2), progesterone (P), sex hormone binding globulin (SHBG), high density-lipoprotein cholesterol (HDL-C), and NET concentrations were measured. Ovulation, delayed ovulation, ovulation inhibition and follicular activity were classified by the analysis of the peripheral serum levels of sex hormones. Ovulation occurred in 7 women in the 0.5 mg group, in 2 women in the 1.5 mg group and in none of the 3.0 mg group. Mean serum SHBG levels were reduced progressively by 6.6% (Group 0.5), 15.5% (Group 1.5) and 23.4% (Group 3.0). There were no significant changes in HDL-C levels in any group. There was a significant correlation of mean serum NET concentrations with dose. The lack of complete inhibition of ovulation in most women in the 1.5 mg and 0.5 mg groups might suggest that the dose of NET required when used as a visiting pill could not be reduced below 3.0 mg.

PIP: In Shanghai, China, 45 25-35 year old women took a daily norethisterone (NET) "visiting pill" (vacation pills) on menstrual cycle days 5-18 as part of a clinical study comparing various doses of NET on ovarian function, sex hormone binding globulin (SHBG), and high density lipoprotein-cholesterol (HDL-C). The aim of the study was to determine the lowest effective dose of the NET visiting pill. Even though some ovarian activity occurred at all 3 doses (0.5, 1.5, and 3 mg), no woman experienced ovulation at 3 mg NET/day during days 5-18 of the cycle. It suppressed ovulation in 11 (73.3%) of the 15 women. Follicular activity occurred in the remaining 4 women. Ovulation occurred in 33% of women taking the 1.5 mg dose and in 66% of those taking the 0.5 mg dose. The higher the NET dose, the greater was the fall in mean serum SHBG levels from control levels (3 mg, 23.4%; 1.5 mg, 15.5%; and 0.5 mg, 6.6%). Both the regression equation and log dose regression equation showed a significant correlation between mean serum NET levels and dose (p .001). HDL-C levels remained basically the same as control levels. Since, at the 1.5 mg dose, ovulation occurred in 5 women and only 5 women experienced complete inhibition, a dose no lower than 3 mg should be used for the NET visiting pill.

A pharmacodynamic and pharmacokinetic study of the Chinese No. 1 pill.

A pharmacodynamic and pharmacokinetic study of the Chinese No. 1 pill, a combined oral contraceptive containing 35 micrograms ethynyloestradiol (EE) and 600 micrograms norethisterone (NET), was performed in 29 women over a period of six months. Blood samples for analysis were taken during a pretreatment cycle, the first and 6th treatment cycles and post-treatment. Minor changes in carbohydrate metabolism occurred and these were particularly noticeable when the incremental areas under the serum concentration-time curves for both glucose and insulin in response to a glucose tolerance test were calculated. No changes occurred in the serum glycosylated haemoglobin levels. The serum concentrations of all the lipids measured (total cholesterol, triglycerides, LDL-C, HDL-C and apolipoproteins AI, AII and B) were significantly increased on treatment as were levels of Factor X, SHBG and caeruloplasmin whereas antithrombin III decreased. In 38 of the 40 treatment cycles, ovulation was suppressed. In one cycle serum oestradiol and progesterone levels showed a typical ovulatory pattern and in another there was evidence of follicular activity without ovulation. Serum EE concentrations showed a similar pattern in both treatment cycles showing that co-administration of NET did not affect EE metabolism. Serum NET levels were higher in the 6th than in the first treatment cycles. On comparing pharmacodynamic and pharmacokinetic parameters, the only statistically significant correlations were between the percentage change in triglycerides and SHBG and serum NET, but not EE concentrations, and between apolipoproteins AI and serum EE.