scholarly journals In vitro degradation of neutral detergent fiber of high-quality tropical forage according to supplementation with different nitrogenous compounds

2009 ◽  
Vol 38 (5) ◽  
pp. 964-971 ◽  
Author(s):  
Karina Zorzi ◽  
Edenio Detmann ◽  
Augusto César de Queiroz ◽  
Mário Fonseca Paulino ◽  
Hilário Cuquetto Mantovani ◽  
...  

The objective of this work was to evaluate the in vitro degradation dynamics of neutral detergent fiber (NDF) of high-quality tropical forage according to supplementation with different nitrogenous compounds sources. A signal grass sample harvested during rainy season was used as basal forage. Casein was used as true protein source and urea:ammonium sulfate (U:AS, 9:1) mixture (urea) as non-protein nitrogen source. The first three supplements were the addition of 0.5, 1.0 and 2.0 mg of casein per mL of final incubation solution. The urea was added to the same amount of protein supplied by equivalent levels 0.5, 1.0 and 2.0 mg/mL of casein. In total, seven treatments were evaluated, including a control (only grass). The supplements were evaluated in a simulated rumen environment by in vitro incubation, and submitted to different incubation times: 0, 3, 6, 9, 12, 24, 36, 48, 72, and 96 hours. The procedure was repeated four times, making a total of four evaluations per incubation time for each treatment. The incubation residues were evaluated for NDF contents and interpreted through a non-linear logistic model. Casein supplementation of 0.5 mg/mL increased the degradation rate of potentially digestible NDF (kpdNDF) by 1.1%. The addition of 1.0 and 2.0 mg/mL of casein provided an inhibitory effect on the estimates of this parameter in comparison to the control (-6.4 and -9.1% respectively). Moreover, the addition of urea, regardless the supplementation level, increased kpdNDF around 7.6%. The ammonia nitrogen (AN) concentration in the medium was increased by supplementation. However, supplementation with urea, even based on the same crude protein levels used with the supplementation with casein, has given higher ammonia nitrogen values in the incubation medium.

2009 ◽  
Vol 38 (10) ◽  
pp. 2021-2030 ◽  
Author(s):  
Isis Lazzarini ◽  
Edenio Detmann ◽  
Cláudia Batista Sampaio ◽  
Mário Fonseca Paulino ◽  
Sebastião de Campos Valadares Filho ◽  
...  

The objective of this study was to evaluate the effects of supplementation with nitrogenous compounds on intake, digestibility, and rumen microbial synthesis in cattle fed low-quality tropical forage. Five crossbred heifers, averaging 209 kg and fitted with ruminal fistulla, were used. The animals were fed ad libitum with signal grass (Brachiaria decumbens Stapf.) hay, which had crude protein (CP) content of 5.08%, as dry matter (DM) basis. The five treatments were defined according to the increasing level of CP in the diet (0, 3, 5, 7, and 9 percentile points above the forage CP level). The supplement consisted of the following nitrogen sources: urea, ammonium sulfate and albumin (4.5:0.5:1.0, respectively). The experiment was carried out according to a 5 × 5 Latin square design, with five experimental periods. The average CP levels in the diets were: 5.28, 8.08, 9.82, 11.87, and 13.63% on DM basis. The intakes of DM, organic matter (OM), and neutral detergent fiber (NDF) showed a quadratic response according to CP levels in the diet, with maximum responses at 10.83%, 10.78%, and 10.37% CP, respectively. The digestibility coefficients of OM and NDF showed a linear-response-plateau response according to CP levels, with the plateau beginning (maximum response) at 7.93% and 7.55% CP, respectively. The average daily concentration of rumen ammonia nitrogen (RAN) was positively related with CP levels. The RAN estimate associated with the maximum DM intake was 15.33 mg/dL. It was observed that intestinal flow of microbial nitrogenous compounds and nitrogen intake became equivalent to each other at 7.13% of CP.


2020 ◽  
Vol 26 (7) ◽  
pp. 532-548 ◽  
Author(s):  
Chunmei Mi ◽  
Bin Ye ◽  
Zhou Gao ◽  
Jinzhi Du ◽  
Ruizhen Li ◽  
...  

Abstract Pre-eclampsia (PE), which results from abnormal placentation, is a primary cause of maternal and neonatal morbidity and mortality. However, the causes of abnormal development of the placenta remain poorly understood. BHLHE40 is a transcriptional repressor in response to hypoxia. Bioinformatics analysis demonstrated that BHLHE40 negatively regulates miR-196a-5p expression, which may decrease miR-196a-5p to target SNX16. Since SNX16 exerts an inhibitory effect on cell migration, it may disrupt trophoblast cell migration in placentation. Therefore, the objective of this study was to explore a possible role of the BHLHE40/miR-196a-5p/SNX16 axis in PE pathogenesis. BHLHE40, miR-196a-5p and SNX16 mRNA and/or protein levels were detected in PE and normal placenta tissues. PE models in vitro and in vivo were constructed by culturing trophoblasts under hypoxia and reducing the uterine perfusion pressure in pregnant C57/BL6N mice, respectively. BHLHE40 and SNX16 were upregulated in PE placenta, while miR-196a-5p was downregulated. Knockdown of BHLHE40 reversed miR-196a-5p expression in trophoblasts under hypoxia, and upregulation of miR-196a-5p inhibited SNX16 expression. As indicated by ChIP assay, BHLHE40 bound to the promoter of the miR-196a-5p gene; luciferase reporter analysis showed that miR-196a-5p could bind to the 3ʹ-untranslated region of SNX16 mRNA. Knockdown of either BHLHE40 or SNX16, or an increase in miR-196a-5p, restored cell viability, migration, invasion and matrix metalloprotein (MMP)-2 and MMP-9 expression under hypoxia. BHLHE40 knockdown also alleviated PE symptoms in pregnant C57/BL6N mice. This study supports involvement of the BHLHE40/miR-196a-5p/SNX16 axis in PE pathogenesis; Proper adjustment of the BHLHE40/miR-196a-5p/SNX16 axis is able to attenuate PE symptoms.


2019 ◽  
Vol 99 (4) ◽  
pp. 741-753
Author(s):  
I.A. Aboagye ◽  
V.S. Baron ◽  
M. Oba ◽  
J. Guyader ◽  
K.A. Beauchemin

In western Canada, short-season corn silage production is increasing due to its potentially high nutritive value. The objective of this study was to determine variability and relationships among nutrient concentration, degradability, and methane (CH4) production of short-season whole-plant corn hybrids harvested before or after light frost (−1.5 °C). Four hybrids, based on their corn heat unit rating (≤2600, CHU rating), were grown in 2 yr in central and southern Alberta (AB) with two field replications. The batch culture and Daisy fermenter techniques were used to characterize degradability and gas production measurements. At both locations, dry matter (DM) concentration was affected by harvest and hybrid (P ≤ 0.02). However, starch and neutral detergent fiber (NDF) concentrations differed (P ≤ 0.01) or tended (P = 0.07) to differ among harvest and hybrid only in central AB. Over both locations and harvest times, CH4production was related negatively to propionate and positively to acetate proportions. In conclusion, harvesting southern AB hybrids after frost increased DM concentration and NDF degradability with no effect on CH4emissions, but the high DM concentration may negatively affect silage quality and animal performance. Harvesting central AB hybrids after frost increased DM and starch concentrations, while reducing CH4emissions but had limited effects on nutrient degradability.


2007 ◽  
Vol 292 (5) ◽  
pp. E1456-E1464 ◽  
Author(s):  
Griselda Irusta ◽  
Fernanda Parborell ◽  
Marta Tesone

Our objective was to study the direct action of a GnRH-I agonist, leuprolide acetate (LA), on ovarian steroidogenesis in preovulatory follicles obtained from equine chorionic gonadotropin (eCG)-treated rats. Previously, we have demonstrated an inhibitory effect of LA on steroidogenesis and follicular development. In this study, we tested the hypothesis that gonadotropin-releasing hormone (GnRH) exerts its negative effect on follicular development by inhibiting thecal cytochrome P-450 C17 (P450C17) α-hydroxylase expression and, consequently, androgen synthesis. Studies were carried out in prepubertal female rats injected with either eCG (control) or eCG plus LA (LA) and killed at different time points. Immunohistochemical studies indicated that LA induced steroidogenic acute regulatory protein (StAR) expression mainly in theca cells of preantral and antral follicles. In addition, serum progesterone levels increased significantly ( P < 0.05), whereas those of androsterone decreased ( P < 0.05) after 8 h of LA treatment. This inhibition caused by LA seemed to be a consequence of the decreased expression of follicular P450C17 α-hydroxylase, as demonstrated by Western blot and RT-PCR techniques. In vitro studies using follicles isolated from 48-h-eCG-treated rats and cultured with LA showed a significant ( P < 0.05) inhibition of FSH-induced androsterone follicular content as well as P450C17 α-hydroxylase protein levels, as determined by Western analysis. However, LA increased StAR protein expression in these follicles without significant changes in P450scc enzyme levels. Taking all these findings into account, we suggest that GnRH-I exerts a direct inhibitory action on gonadotropin-induced follicular development by decreasing the temporal expression of the P450C17 enzyme and, consequently, androgen production, thus reducing the supply of estrogens available to developing follicles.


2021 ◽  
Vol 2021 ◽  
pp. 1-19
Author(s):  
Weiya Cao ◽  
Xueke Liu ◽  
Yinci Zhang ◽  
Amin Li ◽  
Yinghai Xie ◽  
...  

Acquired resistance of hepatocellular carcinoma (HCC) to sorafenib (SFB) is the main reason for the failure of SFB treatment of the cancer. Abnormal activation of the PI3K/AKT/mTOR pathway is important in the acquired resistance of SFB. Therefore, we investigated whether BEZ235 (BEZ) could reverse acquired sorafenib resistance by targeting the PI3K/mTOR pathway. A sorafenib-resistant HCC cell line Huh7R was established. MTT assay, clone formation assay, flow cytometry, and immunofluorescence were used to analyze the effects of BEZ235 alone or combined with sorafenib on cell proliferation, cell cycle, apoptosis, and autophagy of Huh7 and Huh7R cells. The antitumor effect was evaluated in animal models of Huh7R xenografts in vivo. Western blot was used to detect protein levels of the PI3K/AKT/mTOR pathway and related effector molecules. In vitro results showed that the Huh7R had a stronger proliferation ability and antiapoptosis effect than did Huh7, and sorafenib had no inhibitory effect on Huh7R. SFB + BEZ inhibited the activation of the PI3K/AKT/mTOR pathway caused by sorafenib. Moreover, SFB + BEZ inhibited the proliferation and cloning ability, blocked the cell cycle in the G0/G1 phase, and promoted apoptosis in the two cell lines. The autophagy level in Huh7R cells was higher than in Huh7 cells, and BEZ or SFB + BEZ further promoted autophagy in the two cell lines. In vivo, SFB + BEZ inhibited tumor growth by inducing apoptosis and autophagy. We concluded that BEZ235 enhanced the sensitivity of sorafenib through suppressing the PI3K/AKT/mTOR pathway and inducing autophagy. These observations may provide the experimental basis for sorafenib combined with BEZ235 in trial treatment of HCC.


2015 ◽  
Vol 36 (6Supl2) ◽  
pp. 4507
Author(s):  
Luiz Juliano Valério Geron ◽  
Fabiana Gomes da Costa ◽  
Silvia Cristina de Aguiar ◽  
Jocilaine Garcia ◽  
Matheus Gonçalves Ribeiro ◽  
...  

<p>This study evaluated the digestibility of nutrients by, and parameters associated with, <em>in vitro </em>fermentation using different inocula (sheep ruminal fluid and feces) as well as the <em>in vivo </em>digestibility in sheep that were fed rations with 50% concentrate containing either no (0%) residue from the extraction of tamarind pulp (RETP) or 15% RETP. To determine the <em>in vitro </em>digestibility (<em>IV</em>D) of nutrients, two sheep, weighing 40.38 ± 2.10 kg, were used as inoculum donors. To determine the <em>in vivo </em>digestibility of nutrients, we used four sheep and a 3×2 factorial experimental design, with three methods of digestion of nutrients and two experimental rations (0% and 15% RETP). The variables were subjected to analysis of variance and the variables that showed differences at 5% probability were further analyzed using the Tukey test at 5% significance. The <em>IV</em>D using different inocula did not significantly differ (p&gt;0.05) from the <em>in vivo </em>digestibility in sheep for dry matter (DM), organic matter (OM), and neutral detergent fiber (NDF). The different methods for determining nutrient digestibility did not affect (p&gt;0.05) the digestibility of DM, OM, crude protein (CP), NDF, and acid detergent fiber (ADF) in rations with 0% and 15% RETP. However, the <em>IV</em>D of CP for rations containing 0% and 15% RETP incubated with both inocula was lower (p&lt;0.05) than the CP digestibility <em>in vivo</em>. The <em>in vivo </em>digestibility of ADF for rations containing 0% and 15% RETP was higher (P&lt;0.05) than the <em>IV</em>D using sheep ruminal fluid and feces as inocula. The pH values and concentration of ammonia nitrogen (NH3-N) after <em>in vitro </em>incubation for 24 h and the <em>in vivo </em>assay were not different (p&gt;0.05) for the rations containing 0% and 15% RETP, but the pH and NH3-N of both fermented and rumen contents differed (p&lt;0.05) depending on the inocula used and the <em>in vivo </em>assay. In summary, the digestibility of DM, OM, and NDF can be determined by the <em>in vitro </em>fermentation method using the ruminal fluid or feces of sheep as inocula in rations containing 0% or 15% RETP. However, <em>in vitro </em>fermentation is not a suitable method for the determination of pH and NH3-N concentration.</p>


2012 ◽  
Vol 64 (5) ◽  
pp. 1246-1255 ◽  
Author(s):  
J.A.G. Azevêdo ◽  
S.C. Valadares Filho ◽  
D.S. Pina ◽  
E. Detmann ◽  
L.G.R. Pereira ◽  
...  

Fifty-seven by-products were collected from regions throughout Brazil. Chemical composition, in vitro neutral detergent fiber digestibility (IVNDFD), and total digestible nutrients (TDN) were determined with the objective of grouping by-products with similar nutritional characteristics. The by-products belonging to group one (G1) presented the highest content of neutral detergent fiber exclusive of ash and nitrogenous compounds [aNDFom(n)] and lowest energy content, with 42.5% and 38.8% of IVNDFD and TDN, respectively. A new cluster analysis was carried in order to better characterize G2 by-products, six subgroups (SGs) were established (SG1 to SG6). SG1 by-products had the highest and the lowest values for lignin and TDN, respectively. SG2 by-products had the highest aNDFom(n) value, with TDN and IVNDFD values greater than 600 and 700g/kg, respectively, and crude protein (CP) value below 200g/kg in dry matter (DM). Among all the subgroups, SG3 had the highest TDN (772g/kg) and IVNDFD (934g/kg) values and the lowest lignin (23g/kg in DM) value. The ether extract was what most influenced the hierarchical establishment of residual grouping in SG4. SG5 by-products had the highest concentration of non-fibrous carbohydrate. Different from the other subgroups, SG6 by-products had the highest value of available CP.


2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Neuza Mariko Aymoto Hassimotto ◽  
Vanessa Moreira ◽  
Neide Galvão do Nascimento ◽  
Pollyana Cristina Maggio de Castro Souto ◽  
Catarina Teixeira ◽  
...  

Anthocyanins are flavonoids which demonstrated biological activities inin vivoandin vitromodels. Here in the anti-inflammatory properties of an anthocyanin-enriched fraction (AF) extracted from wild mulberry and the cyanidin-3-glucoside (C3G), the most abundant anthocyanin in diet, were studied in two acute inflammation experimental models, in the peritonitis and in the paw oedema assays, both of which were induced by carrageenan (cg) in mice. In each trial, AF and C3G (4 mg/100 g/animal) were orally administered in two distinct protocols: 30 min before and 1 h after cg stimulus. The administration of both AF and C3G suppresses the paw oedema in both administration times (P<0.05). In the peritonitis, AF and C3G reduced the polymorphonuclear leukocytes (PMN) influx in the peritoneal exudates when administered 1 h after cg injection. AF was more efficient reducing the PMN when administered 30 min before cg. Both AF and C3G were found to suppress mRNA as well as protein levels of COX-2 upregulated by cg in both protocols, but the inhibitory effect on PGE2production in the peritoneal exudates was observed when administered 30 min before cg (P<0.05). Our findings suggest that AF and C3G minimize acute inflammation and they present positive contributions as dietary supplements.


2017 ◽  
Vol 52 (12) ◽  
pp. 1268-1275 ◽  
Author(s):  
Leonardo Fiusa de Morais ◽  
João Carlos de Carvalho Almeida ◽  
Delci de Deus Nepomuceno ◽  
Mirton José Frota Morenz ◽  
Bárbara Maria Gomes de Melo ◽  
...  

Abstract: The objective of this work was to evaluate the use of soybean (Glycine max), pigeon pea (Cajanus cajan), and jack bean (Canavalia ensiformis) as urease sources for elephant grass (Pennisetum purpureum) hay ammoniated with urea. The experimental design was completely randomized in a double factorial arrangement with one additional treatment: 4 urease source levels x 3 urease sources + 1 control. Chemical-bromatological analyses and carbohydrate fractionation were performed in the hay, and cumulative gas production in vitro was determined. There were interactions between urease level and source for neutral detergent fiber and acid detergent fiber, in which 1 and 2% jack bean lowered acid detergent fiber values, and 2% jack bean and 3% soybean reduced lignin content. The addition of milled legume grains reduces fiber components and increases non protein nitrogen content in elephant grass hay ammoniated with urea. Adding 4% milled soybean increases gas production in the soluble fraction.


Endocrinology ◽  
2015 ◽  
Vol 156 (11) ◽  
pp. 4094-4104 ◽  
Author(s):  
Jan Schovanek ◽  
Petra Bullova ◽  
Yasin Tayem ◽  
Alessio Giubellino ◽  
Robert Wesley ◽  
...  

Metastatic pheochromocytoma continues to be an incurable disease, and treatment with conventional cytotoxic chemotherapy offers limited efficacy. In the present study, we evaluated a novel topoisomerase I inhibitor, LMP-400, as a potential treatment for this devastating disease. We found a high expression of topoisomerase I in human metastatic pheochromocytoma, providing a basis for the evaluation of a topoisomerase 1 inhibitor as a therapeutic strategy. LMP-400 inhibited the cell growth of established mouse pheochromocytoma cell lines and primary human tumor tissue cultures. In a study performed in athymic female mice, LMP-400 demonstrated a significant inhibitory effect on tumor growth with two drug administration regimens. Furthermore, low doses of LMP-400 decreased the protein levels of hypoxia-inducible factor 1 (HIF-1α), one of a family of factors studied as potential metastatic drivers in these tumors. The HIF-1α decrease resulted in changes in the mRNA levels of HIF-1 transcriptional targets. In vitro, LMP-400 showed an increase in the growth-inhibitory effects in combination with other chemotherapeutic drugs that are currently used for the treatment of pheochromocytoma. We conclude that LMP-400 has promising antitumor activity in preclinical models of metastatic pheochromocytoma and its use should be considered in future clinical trials.


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