RC Treatment Research Articles

Ginsenoside Rc alleviates osteoporosis by the TGF-β/Smad signaling pathway.

Osteoporosis is a common chronic bone disorder in postmenopausal women. Ginsenosides are primary active components in ginseng and the effects of various ginsenoside variants in osteoporosis treatment have been widely revealed. We planned to explore the impact of ginsenoside Rc on bone resorption in an osteoporosis rat model. We used ovariectomized rats to assess the potential impact of ginsenoside Rc on osteoporosis. μ-CT was implemented for analyzing the microstructure of the distal left femur in rats. H&E staining together with Masson staining were applied for bone histomorphometry evaluation. ELISA kits were implemented to detect serum concentrations of TRACP-5b, OCN, CTX, as well as PINP. Ginsenoside Rc treatment lessened the serum levels of TRACP-5b as well as CTX, while increasing serum levels of OCN, and PINP of OVX rats. Moreover, we found that ginsenoside Rc contributed to the synthesis of type I collagen via increasing Col1a1 and Col1a2 levels in femur tissues of ovariectomized rats. Our findings also revealed that ginsenoside Rc activated the TGF-β/Smad pathway by increasing TGF-β as well as phosphorylated Smad2/3 protein levels. Ginsenoside Rc alleviates osteoporosis in rats through promoting the TGF-β/Smad pathway.

Rapid composting of groundnut residues through novel microbial consortium: Evaluating maturity, stability, and microbial activity

A laboratory pot experiment (Experiment 1) was conducted to determine the optimal ratio of groundnut haulm and shell as composting substrates. The aim was to identify the most effective combination for rapid decomposition under in vivo conditions. The experiment was carried out in 2022, from May to July, using a completely randomized design (CRD) with 6 treatments that were replicated 4 times. The treatment combinations in the pot experiment were as follows: T1: haulm + shell (1:1), T2: haulm + shell (2:1), T3: haulm + shell (3:1), T4: T1 + Cn, T5: T2 + Cn, and T6: T3 + Cn. Here, Cn refers to the cellulose-degrading efficient microbial consortium containing bacterial strains Priestia megaterium DBJ6, Micrococcus yunnanensis DMB9, and fungal strains Aspergillus foetidus DAJ2, Trichoderma atrobrunnium DTJ4, and Phanerochaete chrysosporium CBS129.27. Based on the results (results of chemical and biological properties) of the pot experiment, treatment T6 was considered the best treatment (the C/N ratio was 14.36 ± 0.444 after three months of decomposition) for further study under in vivo conditions. The in vivo experiment (Experiment 2) was conducted at the Jaguli Instructional Farm of Bidhan Chandra Krishi Viswavidyalaya, West Bengal, India, during the months of August to October in both 2022 and 2023 with two treatments: the normal composting treatment (NC) and the rapid composting treatment (RC). The findings of Experiment 2 showed that organic substrates degraded more quickly (within 90 days) during rapid composting, as evidenced by a decrease in the C/N ratio to below 15 (14.32 and 13.97 on day 90 in 2022 and 2023, respectively). Whereas, normal composting required more than 90 days to achieve a C/N ratio below 20 (23.80 and 23.15 on day 90 in 2022 and 2023, respectively). The RC treatment also showed a higher microbial population and enzyme activity compared to the NC treatment. Therefore, according to the results of this experiment, it can be concluded that the inoculation of the microbial consortium was more beneficial for accelerating the composting process under the same composting circumstances and substrate ratio.

Fungal pretreatment of citrus waste improves the hydrolysis and acidogenesis of the organic fraction of urban solids wastes

This work shows the increase in hydrolysis and acidogenesis of the organic fraction of urban solid waste (OFUSW) by the action of fungal hydrolytic enzymes obtained by solid-state fermentation (SSF) with Aspergillus niger C25B28 on citrus wastes (CW) without sterilization or the addition of nutrients. To quantify the hydrolysis and acidogenesis of the organic fraction of urban solid waste, three treatments were studied in triplicate: OFUSW plus 10 % fresh CW (RC treatment), and OFUSW plus 10 % citrus wastes pretreated by SSF (RCA treatment). The mixtures were packed into an anaerobic hydrolytic leaching bed reactor. The results showed that SSF decreased the limonene concentration by 97 %, removing 20 % of fiber and 46 % of fat from fresh CW. Pectinases (37 U/g DM [dry matter]), cellulases (33 U/g DM), xylanases (98 U/gDM), and proteases (0.2 U/gDM) were produced. Significant differences were observed between the different treatments: the greatest reduction in total solids (24 %), the highest removal efficiency of organic matter as chemical oxygen demand (COD), and the reduction of fats (23 % and 28 %, respectively), was achieved in RCA, compared to those obtained in the reactors with OFUSW and ReC without pretreatment. The production rate of volatile fatty acids (VFA) was 1.4 gVFA/Ld for RCA, 4.5 times higher than in the reactors without pretreatment.

Potato and soil 15N recoveries from different labelled forage root and shoot

An improved understanding of the contribution of a preceding forage crop to a subsequent potato crop can improve nitrogen (N) utilization in potato production. This study used two rotation experiments to estimate the N contribution from labelled shoot and root of red clover (RC, Trifolium pratense), timothy (T, Phleum pratense) and a red clover/timothy mixture (M) to a subsequent potato crop using microplots in the field. Forage crops were grown with 14NH414NO3 and 15NH415NO3 (98 atom %). The residue exchange technique was used to compare residue treatments of (i) whole plant labelled; (ii) labelled shoot only; and (iii) labelled root only in Experiment 1, and residue treatments of (i) whole plant labelled; (ii) labelled shoot/unlabelled root; and (iii) labelled root/unlabelled shoot in Experiment 2. Averaged across forage treatments, recoverable root biomass represented 64 and 37% of total forage biomass, and the total 15N recovery from labelled roots was 52 and 62% of the total 15N recovery from shoots, in Experiments 1 and 2, respectively. Therefore, forage roots represented a substantial source of N for the subsequent crop. However, less than 5% of the 15N from crop residues was recovered in the potato vines plus tubers, and most of the 15N was recovered in the soil, regardless of the forage or residue treatments. Potato tuber and vine dry matter was greater for the RC than the T treatment for all residue treatments, a finding attributed to greater potato N accumulation for the RC treatment. It is therefore important to consider the contribution of forage roots when studying N cycling in potato systems. Potato N requirements were satisfied more by soil-derived N rather than from fall incorporated forage residues.

Ginsenoside Rc attenuates DSS-induced ulcerative colitis, intestinal inflammatory, and barrier function by activating the farnesoid X receptor.

Objectives: Farnesoid X receptor (FXR) activation is involved in ameliorating inflammatory bowel disease (IBD), such as ulcerative colitis (UC), and inflammatory regulation may be involved in its mechanism. Ginsenoside Rc (Rc) is a major component of Panax ginseng, and it plays an excellent role in the anti-inflammatory processes. Our aim is to explore the alleviative effect of Rc on dextran sulfate sodium (DSS)-induced inflammation and deficiencies in barrier function based on FXR signaling. Materials and Methods: In vitro, we treated human intestinal epithelial cell lines (LS174T) with LPS to explore the anti-inflammatory effect of Rc supplementation. In vivo, a DSS-induced IBD mice model was established, and the changes in inflammatory and barrier function in colons after Rc treatment were measured using the disease activity index (DAI), hematoxylin and eosin (H&E) staining, immunofluorescence, ELISA, and qPCR. Molecular docking analysis, luciferase reporter gene assay, and qPCR were then used to analyze the binding targets of Rc. DSS-induced FXR-knockout (FXR-/-) mice were used for further validation. Results: Rc significantly recovered the abnormal levels of inflammation indexes (TNF-α, IL-6, IL-1β, and NF-KB) induced by LPS in LS174T. DSS-induced C57BL/6 mice exhibited a significantly decreased body weight and elevated DAI, as well as a decrease in colon weight and length. Increased inflammatory markers (TNF-α, IL-6, IL-1β, ICAM1, NF-KB, F4/80, and CD11b displayed an increased expression) and damaged barrier function (Claudin-1, occludin, and ZO-1 displayed a decreased expression) were observed in DSS-induced C57BL/6 mice. Nevertheless, supplementation with Rc mitigated the increased inflammatory and damaged barrier function associated with DSS. Further evaluation revealed an activation of FXR signaling in Rc-treated LS174T, with FXR, BSEP, and SHP found to be upregulated. Furthermore, molecular docking indicated that there is a clear interaction between Rc and FXR, while Rc activated transcriptional expression of FXR in luciferase reporter gene assay. However, these reversal abilities of Rc were not observed in DSS-induced FXR-/- mice. Conclusion: Our findings suggest that Rc may ameliorate inflammation and barrier function in the intestine, which in turn leads to the attenuation of DSS-induced UC, in which Rc may potentially activate FXR signaling to protect the intestines from DSS-induced injury.

Ginsenoside Rc, as an FXR activator, alleviates acetaminophen-induced hepatotoxicity via relieving inflammation and oxidative stress.

Acetaminophen (APAP) intake leads to excessive NAPQI deposition, stimulating inflammatory and oxidative stress and causing fatal liver injury. However, the detailed molecular mechanism involved is unknown, and effective therapeutic approaches remain insufficient. In this study, we discovered that treatment with ginsenoside Rc can prevent the inflammatory response caused by APAP and oxidative stress in mouse primary hepatocytes (MPHs), along with the corresponding changes in related genes. Additionally, Ginsenoside Rc effectively alleviates APAP-induced cellular apoptosis and NAPQI accumulation in MPHs. In vivo, Ginsenoside Rc administration remarkably attenuates APAP-induced hepatotoxicity, repairing liver damage and improving survival. Moreover, Ginsenoside Rc treatment modulates genes involved in APAP metabolism, leading to a decrease in NAPQI and resulting in the alleviation of fatal oxidative stress and inflammatory response after APAP exposure, along with the expression of their related indicators. Furthermore, our RNA-seq and molecular docking analysis implies that FXR expression and FXR transcriptional activity are stimulated by Ginsenoside Rc treatment. Notably, due to the lack of FXR in mice and MPHs, ginsenoside Rc can no longer play its original protective role against hepatotoxicity and cell damage caused by APAP, and it is difficult to improve the corresponding survival rate and prevent hepatic apoptosis, NAPQI generation, fatal oxidative stress, and the inflammatory response induced by APAP and the expression of related genes. In summary, our results indicate that Ginsenoside Rc could act as an effective FXR activator and effectively regulate FXR-induced antioxidant stress and eliminate inflammation while also having an anti-apoptotic function.

The underlying rationality of Chinese medicine herb pair Coptis chinensis and Dolomiaea souliei: From the perspective of metabolomics and intestinal function

Ethnopharmacological relevanceThe combination of Coptis chinensis (RC) and Dolomiaea souliei (VR) has long been used as a classic herb pair for the treatment of gastrointestinal diseases, but the underlying mechanisms remain unknown. Materials and methodsIn this study, the rationality of evidence-based RC and VR combination was explored from the perspective of metabolism, gut microbiota and gastrointestinal function. ResultsAfter 5 weeks treatment, VR extracts (700 mg/kg) and RC alkaloids (800 mg/kg) showed no toxic effect on mice. However, RC administration significantly decreased the body weight of mice. Gastric emptying, gastrointestinal motility function and the absorption of FITC dextran were retarded in the mice of RC group, taking RC along with low dose VR (RC-VRL) and high dose VR (RC-VRH) reversed the impaired gastrointestinal function caused by RC. RC administration significantly increased villus height/crypt depth value. Notably, VR administration increased the number of crypts in mice ileum and reduced villus height/crypt depth value in VR and RC combination group. RC treatment significantly increased the expression of occludin compared to NC group; RC-VRL treatment reversed this tendency. While, VR administration increased ZO1 expression by 99.4% compared to NC mice. As for gut microbiota, RC gavage decreased the gut microbiota diversity, but gut microbiota in VR group was similar to NC group, and VR and RC combination increased gut microbiota diversity. RC administration obviously increased the proportion of Akkermansia muciniphila, Bacteroides thetaiotaomicron, Parabacteroides distasonis, and Escherichia coli, compared to NC mice. VR treatment increased the richness of Bacteroides thetaiotaomicron, Parabacteroides distasonis. RC-VRL and RC-VRH treatment dose-dependently increased the richness of Rikenellaceae RC9, Lactobacillus, and decreased the abundance of Psychrobacter, Bacteroides and Ruminococcus in mice. Serum metabolomic analysis revealed that RC gavage significantly down regulated 76 metabolites and up regulated 31 metabolites. VR treatment significantly down regulated 30 metabolites and up regulated 12 metabolites. Weight loss caused by RC may attribute to the elevated methylxanthine level in mice. The potential adverse effects caused by high dose RC intake may partially alleviate by high serum contents of adenosine, inosine and urolithin A resulted from VR coadministration. ConclusionVR may alleviate RC caused “fluid retention” via normalizing gastrointestinal function, gut microbiota and modulating the perturbed metabolism.

Ginsenosides Rc, as a novel SIRT6 activator, protects mice against high fat diet induced NAFLD

BackgroundHepatic lipid disorder impaired mitochondrial homeostasis and intracellular redox balance, triggering development of non-alcohol fatty liver disease (NAFLD), while effective therapeutic approach remains inadequate. Ginsenosides Rc has been reported to maintain glucose balance in adipose tissue, while its role in regulating lipid metabolism remain vacant. Thus, we investigated the function and mechanism of ginsenosides Rc in defending high fat diet (HFD)-induced NAFLD. MethodsMice primary hepatocytes (MPHs) challenged with oleic acid & palmitic acid were used to test the effects of ginsenosides Rc on intracellular lipid metabolism. RNAseq and molecular docking study were performed to explore potential targets of ginsenosides Rc in defending lipid deposition. Wild type and liver specific sirtuin 6 (SIRT6, 50721) deficient mice on HFD for 12 weeks were subjected to different dose of ginsenosides Rc to determine the function and detailed mechanism in vivo. ResultsWe identified ginsenosides Rc as a novel SIRT6 activator via increasing its expression and deacetylase activity. Ginsenosides Rc defends OA&PA-induced lipid deposition in MPHs and protects mice against HFD-induced metabolic disorder in dosage dependent manner. Ginsenosides Rc (20mg/kg) injection improved glucose intolerance, insulin resistance, oxidative stress and inflammation response in HFD mice. Ginsenosides Rc treatment accelerates peroxisome proliferator activated receptor alpha (PPAR-α, 19013)-mediated fatty acid oxidation in vivo and in vitro. Hepatic specific SIRT6 deletion abolished ginsenoside Rc-derived protective effects against HFD-induced NAFLD. ConclusionGinsenosides Rc protects mice against HFD-induced hepatosteatosis by improving PPAR-α-mediated fatty acid oxidation and antioxidant capacity in a SIRT6 dependent manner, and providing a promising strategy for NAFLD.

Agroecological practices for organic lettuce: effects on yield, nitrogen status and nitrogen utilisation efficiency

ABSTRACTClimate change tends to intensify extreme weather events worldwide, negatively affecting agriculture and threatening global food security. More resilient agroecosystems are required to assure yield stability. In this research, which was carried out in a Mediterranean environment in 2013–2015, the response of organic lettuce to cover crop management and organic fertilisation, in terms of yield, nitrogen status and efficiency, was investigated. A vetch cover crop was introduced before the lettuce and was terminated by the no-till roller crimper strategy (RC), which was compared with incorporation of the vetch into the soil as green manure (GM) and with a no-vetch control (No_V). Two organic fertilisers derived from waste recycling, anaerobic digestate (AD) and municipal solid waste compost (MSW), were compared with a commercial organic fertiliser (ORG) and with a control with no fertiliser (No_F). The RC treatment resulted in high lettuce yields in both years, whereas GM showed intermediate yield values (significant differences in 2014). Crop N status reflected yield, with improved values in RC plots. At harvest, particularly in 2014, the use of vetch and fertilisers increased soil mineral nitrogen concentrations compared with those in No_V and No_F treatments, though the differences were not statistically significant. Lettuce N use efficiency (NUE) had an inverse trend to that of the yield; the lower the yield the higher the NUE. The RC in combination with the use of organic fertilisers assured a satisfactory response to the variability in weather, in terms of both yield stability and preservation of soil fertility.

Effects of straw returning and feeding on greenhouse gas emissions from integrated rice-crayfish farming in Jianghan Plain, China.

Great efforts have been devoted to assessing the effects of straw managements on greenhouse gas (GHG) emissions, global warming potential (GWP), and net economic budget in rice monoculture (RM). However, few studies have evaluated the effects of straw managements on GHG emissions and net ecosystem economic budget (NEEB) in integrated rice-crayfish farming (RC). Here, a randomized block field experiment was performed to comprehensively evaluate the effects of aquatic breeding practices (feeding or no feeding of forage) and straw managements (rice straw returning or removal) on soil NH4+-N and NO-3-N contents, redox potential (Eh), CH4 and N2O emissions, GWP, and NEEB of fluvo-aquic paddy soil in a rice-crayfish co-culture system in Jianghan Plain of China. We also compared the differences in CH4 and N2O emissions, GWP, and NEEB between RM and RC. Straw returning significantly increased CH4 and N2O emissions by 34.9-46.1% and 6.2-23.1% respectively compared with straw removal. Feeding of forage decreased CH4 emissions by 13.9-18.7% but enhanced N2O emissions by 24.4-33.2% relative to no feeding. Compared with RM treatment, RC treatment decreased CH4 emissions by 18.1-19.6% but increased N2O emissions by 16.8-21.0%. Moreover, RC treatment decreased GWP by 16.8-22.0% while increased NEEB by 26.9-75.6% relative to RM treatment, suggesting that the RC model may be a promising option for mitigating GWP and increasing economic benefits of paddy fields. However, the RC model resulted in a lower grain yield compared with the RM model, indicating that more efforts are needed to simultaneously increase grain yield and NEEB and decrease GWP under RC model.

The Effects of Rice Straw and Biochar Applications on the Microbial Community in a Soil with a History of Continuous Tomato Planting History

Soil microbial abundance and diversity change constantly in continuous cropping systems, resulting in the prevalence of soil-borne pathogens and a decline in crop yield in solar greenhouses. To investigate the effects of rice straw and biochar on soil microbial abundance and diversity in soils with a history of continuous planting, three treatments were examined: mixed rice straw and biochar addition (RC), rice straw addition (R), and biochar addition (C). The amount of C added in each treatment group was 3.78 g kg−1 soil. Soil without rice straw and biochar addition was treated as a control (CK). Results showed that RC treatment significantly increased soil pH, available nitrogen (AN), available phosphorus (AP), and potassium (AK) by 40.3%, 157.2%, and 24.2%, respectively, as compared to the CK soil. The amount of soil labile organic carbon (LOC), including readily oxidizable organic carbon (ROC), dissolved organic carbon (DOC), and light fraction organic carbon (LFOC), was significantly greater in the RC, R, and C treatment groups as compared to CK soil. LOC levels with RC treatment were higher than with the other treatments. Both rice straw and biochar addition significantly increased bacterial and total microbial abundance, whereas rice straw but not biochar addition improved soil microbial carbon metabolism and diversity. Thus, the significant effects of rice straw and biochar on soil microbial carbon metabolism and diversity were attributed to the quantity of DOC in the treatments. Therefore, our results indicated that soil microbial diversity is directly associated with DOC. Based on the results of this study, mixed rice straw and biochar addition, rather than their application individually, might be key to restoring degraded soil.

Effects of conditioners (single-layer, double-layer and retention-conditioner) on the growth performance, meat quality and intestinal morphology of growing and finishing pigs

This experiment was conducted to investigate the effects of feed conditioners (single-layer, double-layer and retention-conditioner) on the growth performance, meat quality and intestinal morphology of pigs throughout the growing to finishing phase. A total of 96 growing pigs ((28.70±3.20) kg) were selected and randomized into three treatment groups with four replicates per group. Eight pigs were used per replicate for the 17-week feeding trial. The grower diet was given at 0 to 6 weeks and a finisher diet was given at 6 to 17 weeks. The treatments were as follows: SC diet (control; single-layer conditioning), DC diet (double-layer conditioning), and RC diet (retention-conditioning). Starch gelatinization was significantly higher (P<0.05) in the RC treatment than in the SC treatment, however, there was no significant difference in the starch gelatinization between the DC group and the RC group. In the growing phase, the feed to gain index (F:G) was significantly lower (P<0.05) in the RC group than in the SC and DC groups. Between growing and finishing, the F:G was the lowest (P<0.05) in the SC group compared to the RC or DC group. Drip loss, a measurement of meat quality, was significantly lower (P<0.05) in longissimus dorsi tissue collected from pigs fed the RC diet than in tissues collected from pigs fed the SC diet. The intestinal quality of the duodenum and jejunum tissues showed a significant increase (P<0.05) in the crypt depth and villus height in the RC group compared to the SC- or DC-treated pigs. These results demonstrated that the retention-conditioner treatment decreased the F:G in growing pigs, improved intestinal morphology and enhanced the meat quality in the finishing pigs. However, the retention-conditioner treatment had a negative impact on growth performance in the finishing pigs.

Ginsenoside Rc protects against UVB‑induced photooxidative damage in epidermal keratinocytes.

Ginsenoside Rc (Rc) is a major ginsenoside isolated from Panaxginseng, and has exhibited pharmacological effects on skin cells. The present study aimed to investigate the putative skin‑protective properties of Rc, including its anti‑photoaging and barrier function‑protective effects, in human HaCaT keratinocytes exposed to UVB radiation. The protective properties of Rc were evaluated through the assessment of keratinocyte viability, reactive oxygen species (ROS) production, total glutathione (GSH) and superoxide dismutase (SOD) activity, caspase‑14, matrix metalloproteinase (MMP)‑2 and ‑9 activity, and MMP‑2, MMP‑9 and filament aggregating protein (filaggrin) expression following UVB irradiation. Treatment with Rc was revealed to prevent the UVB‑induced increase in ROS production and pro‑MMP‑2 and ‑9 levels in HaCaT keratinocytes. In addition, treatment with Rc resulted in enriched GSH contents and enhanced SOD activity following exposure to UVB radiation. Furthermore, Rc treatment enhanced caspase‑14 activity and counteracted the UVB‑induced downregulation in filaggrin expression. However, no significant difference was identified between Rc‑treated and normal groups in terms of keratinocyte viability, regardless of exposure to radiation. The present findings suggested that Rc may exert anti‑photoaging and barrier function‑protective effects in keratinocytes, and thus protect the skin against photooxidative stress induced by exposure to UV radiation.

Quantification of Hemoglobin and Myoglobin in Pork Muscle: Effect of Rinse&Chill Technology on Blood Removal

ObjectivesThe objective of this study was to use ion exchange chromatography to measure hemoglobin (Hb) and myoglobin (Mb) content in ground and salted pork lean obtained from early post-mortem sow carcasses treated with RinseC balance: glucose, polyphosphates, glycerine, and maltose). Sows were electrically stunned (550 V) prior to exsanguination. Six sows were used as the conventional treatment and 6 sows received the RC process. At 30 to 60 min post-mortem, the Boston butt and picnic shoulder were deboned, ground (9.5 mm diameter plate) and mixed with 1% NaCl (w/w). Samples (∼100 g) were vacuum-packaged and stored at –80°C until analysis. After thawing, samples were frozen with liquid nitrogen and ground into a fine powder for extraction and determination of total heme, Mb, and Hb. Total heme was extracted with acid acetone and determined spectrophotometrically (e₆₄₀nm = 4.80 mM–¹cm–¹). Mb and Hb were extracted with 0.01 M PBS buffer (pH 7.4) and after exchange into a 0.01 M Tris buffer (pH 8.6), the heme pigments were separated by ion exchange chromatography in a diethylaminoethyl cellulose (DE52 resin, 2 g) column. Mb was eluted with 0.05 M Tris buffer (pH 8.0) and Hb with 0.5 M NaCl solution. The amount of Mb and Hb in the corresponding eluates was determined spectrophotometrically (e₄₁₈nm = 125 mM–¹cm–¹ for Mb and e₄₁₄nm = 128 mM–¹cm–¹ for Hb). ESI-MS was used to measure the mass of polypeptides in the Mb and Hb fractions. Total heme, Mb, and Hb content in muscle extracts from the 2 treatments were compared using an unpaired student t test. ResultsESI-MS analysis of the Mb fraction indicated a mass of 16,954 Da which was consistent with the mass of pig Mb without its heme moiety. The Hb fraction had masses of 15,040 and 16,036 Da which was consistent with the mass of the ɑ and β chain of pig Hb, respectively, without their heme moiety. These results indicated that the chromatographic separation was satisfactory. Heme and Mb content in the muscle extracts were statistically similar, when comparing conventional to RC process. The corresponding means for heme content were 187.5 and 209.2 μmol/kg muscle and for Mb, 151.9 and 168.1 μmol/kg muscle. Hb content in the RC treatment was significantly lower, 39.6% on average, than that for the conventional treatment. The percentages of Mb and Hb by weight (relative to each other) in the conventional treatment were 80.9 and 19.1%, respectively. For the RC treatment, Mb was 86.4% and Hb was 13.6%. ConclusionThe ion exchange chromatography method presented here allowed the measurement of Mb and Hb in pork lean with a satisfactory recovery compared to the total heme determination. Using Hb content in muscle extracts as a measure of blood content suggested that the RC method removed 40% more blood from the muscle compared to the conventional method, leaving still about 60% of the blood in the muscle of RC-treated pigs. The possibility to quantify Hb and Mb separately offers a method to quantitatively assess blood content in muscle.

Quantification of Hemoglobin and Myoglobin in Pork Muscle: Effect of Rinse&amp;Chill Technology on Blood Removal

ObjectivesThe objective of this study was to use ion exchange chromatography to measure hemoglobin (Hb) and myoglobin (Mb) content in ground and salted pork lean obtained from early post-mortem sow carcasses treated with RinseC balance: glucose, polyphosphates, glycerine, and maltose). Sows were electrically stunned (550 V) prior to exsanguination. Six sows were used as the conventional treatment and 6 sows received the RC process. At 30 to 60 min post-mortem, the Boston butt and picnic shoulder were deboned, ground (9.5 mm diameter plate) and mixed with 1% NaCl (w/w). Samples (∼100 g) were vacuum-packaged and stored at –80°C until analysis. After thawing, samples were frozen with liquid nitrogen and ground into a fine powder for extraction and determination of total heme, Mb, and Hb. Total heme was extracted with acid acetone and determined spectrophotometrically (e₆₄₀nm = 4.80 mM–¹cm–¹). Mb and Hb were extracted with 0.01 M PBS buffer (pH 7.4) and after exchange into a 0.01 M Tris buffer (pH 8.6), the heme pigments were separated by ion exchange chromatography in a diethylaminoethyl cellulose (DE52 resin, 2 g) column. Mb was eluted with 0.05 M Tris buffer (pH 8.0) and Hb with 0.5 M NaCl solution. The amount of Mb and Hb in the corresponding eluates was determined spectrophotometrically (e₄₁₈nm = 125 mM–¹cm–¹ for Mb and e₄₁₄nm = 128 mM–¹cm–¹ for Hb). ESI-MS was used to measure the mass of polypeptides in the Mb and Hb fractions. Total heme, Mb, and Hb content in muscle extracts from the 2 treatments were compared using an unpaired student t test. ResultsESI-MS analysis of the Mb fraction indicated a mass of 16,954 Da which was consistent with the mass of pig Mb without its heme moiety. The Hb fraction had masses of 15,040 and 16,036 Da which was consistent with the mass of the ɑ and β chain of pig Hb, respectively, without their heme moiety. These results indicated that the chromatographic separation was satisfactory. Heme and Mb content in the muscle extracts were statistically similar, when comparing conventional to RC process. The corresponding means for heme content were 187.5 and 209.2 μmol/kg muscle and for Mb, 151.9 and 168.1 μmol/kg muscle. Hb content in the RC treatment was significantly lower, 39.6% on average, than that for the conventional treatment. The percentages of Mb and Hb by weight (relative to each other) in the conventional treatment were 80.9 and 19.1%, respectively. For the RC treatment, Mb was 86.4% and Hb was 13.6%. ConclusionThe ion exchange chromatography method presented here allowed the measurement of Mb and Hb in pork lean with a satisfactory recovery compared to the total heme determination. Using Hb content in muscle extracts as a measure of blood content suggested that the RC method removed 40% more blood from the muscle compared to the conventional method, leaving still about 60% of the blood in the muscle of RC-treated pigs. The possibility to quantify Hb and Mb separately offers a method to quantitatively assess blood content in muscle.

Crop residue management and fertilization effects on soil organic matter and associated biological properties.

Returning crop residue may result in nutrient reduction in soil in the first few years. A two-year field experiment was conducted to assess whether this negative effect is alleviated by improved crop residue management (CRM). Nine treatments (3 CRM and 3N fertilizer rates) were used. The CRM treatments were (1) R0: 100% of the N using mineral fertilizer with no crop residues return; (2) R: crop residue plus mineral fertilizer as for the R0; and (3) Rc: crop residue plus 83% of the N using mineral and 17% manure fertilizer. Each CRM received N fertilizer rates at 270, 360, and 450kgNha(-1) year(-1). At the end of the experiment, soil NO3-N was reduced by 33% from the R relative to the R0 treatment, while the Rc treatment resulted in a 21 to 44% increase in occluded particulate organic C and N, and 80°C extracted dissolved organic N, 19 to 32% increase in microbial biomass C and protease activity, and higher monounsaturated phospholipid fatty acid (PLFA):saturated PLFA ratio from stimulating growth of indigenous bacteria when compared with the R treatment. Principal component analysis showed that the Biolog and PLFA profiles in the three CRM treatments were different from each other. Overall, these properties were not influenced by the used N fertilizer rates. Our results indicated that application of 17% of the total N using manure in a field with crop residues return was effective for improving potential plant N availability and labile soil organic matter, primarily due to a shift in the dominant microorganisms.

Induction of Apoptosis by Methanolic Extract of Rubia Cordifolia Linn in HEp-2 Cell Line is Mediated by Reactive Oxygen Species

Rubia cordifolia Linn, which belongs to the Rubiaceae family, is a well-known herb used in Ayurvedic medicine. In the present study, we investigated the influence of a methanolic extract (RC) on the induction of apoptosis in HEp-2 (human laryngeal carcinoma) cell line, as evidenced by cytotoxicity, morphological changes and modification in the levels of pro-oxidants. Inhibition of cell proliferation and lactate dehydrogenase (LDH) release increased in a time and dose-dependent manner. Further, reduced glutathione (GSH), glutathione transferase (GST) and protein levels decreased and lipid peroxidation increased significantly on RC treatment in a dose dependent manner when compared to controls. Based on the results we determined the optimal dose as 30 mg/ ml and the apoptotic effect of RC extract (30 mg/ml) on HEp-2 cells was confirmed by fluorescent microscopy and transmission electron microscopy (TEM) based on morphological and ultrastructural changes. RC extract suppressed the proliferation of HEp-2 oral cancer cells inducing apoptotic cell death in vitro. These results point to potential of RC extract as an agent for the treatment of laryngeal squamous cell carcinoma.

Measurement of the Intracellular pH in Human Stomach Cells: A Novel Approach To Evaluate the Gastric Acid Secretory Potential of Coffee Beverages

As the consumption of coffee beverages sometimes is reported to cause gastric irritation, for which an increased stomach acid secretion is one of the promoting factors, different processing technologies such as steam-treatment have been developed to reduce putative stomach irritating compounds. There is evidence-based data neither on the effect of detailed processing variations nor on individual coffee components affecting the proton secretory activity (PSA). This work aimed at developing a screening model suitable for investigating the effects of commercial coffee beverages and components thereof on human parietal cells. Human gastric cancer cells (HGT-1) were treated with reconstituted freeze-dried coffee beverages prepared from customary coffee products such as regular coffee (RC, n = 4), mild bean coffee (MBC, n = 5), stomach friendly coffee (SFC, n = 4), and SFC decaffeinated (SFCD, n = 3). PSA was analyzed by flow cytometry using the pH-sensitive dye SNARF-AM. Treatment of the cells with MBC did not result in a PSA different from RC treatment (p <or= 0.07), whereas cells treated with SFC (p <or= 0.04) or SFCD (p <or= 0.03) showed a significantly lower PSA than those treated with RC. Quantitative and principle component analysis of putative stomach irritating compounds revealed significantly reduced contents of (beta)N-alkanoyl-5-hydroxytryptamides, caffeine, N-methylpyridinium, and catechol in SFCD compared to RC. However, none of these compounds seem to act as the sole key bioactive reducing the PSA of SFCD, since their contents in MBC and SFC samples were not different from those in RC samples, although the PSA of these beverages was significantly lower than that of reconstituted freeze-dried RC beverage.

Effect Of Rhodiola Crenulata And Cordyceps Sinensis-based Oral Supplementation On Exercise Stress Test In Altitude

PURPOSE: To determine the effect to Rhodiola Crenulata and Cordyceps Sinensis-Based Oral Supplementation on muscle oxygenation after exercise at sea-level and altitude. METHODS: Eighteen male subjects were weight-matched and evenly divided into 2 groups: Oral Rhodiola Crenulata and Cordyceps Sinensis (RC) supplementation and Placebo groups. A standardized 3-min step test was performed at sea-level and altitude (2200 meters). Arterial oxygen saturation (SaO2) and muscle tissue oxygenation (StO2) were measured before and after exercise. Running performance was determined by graded Bruce‘s protocol. RESULTS: RC supplementation did not alter maximal oxygen consumption and heart rate recovery on 3-min step test. Blood lactate recoveries following both step test and all-out running was not altered by RC treatment. However, SaO2 was lower in the RC group than that in placebo group. Conversely, total hemoglobin concentration and percent oxygen saturation in muscle tissue were significantly greater in the RC group than those in the placebo group. CONCLUSIONS: Despites RC supplementation has no significant acute effect on exercise performance, this supplementation resulted in a greater oxygen delivery to skeletal muscle.

Effect of feeding fresh or conditioned red clover on milk fatty acids and nitrogen utilization in lactating dairy cows

Polyphenol oxidase (PPO) in conditioned red clover (ensiled or cut and crushed) reduces both proteolysis and lipolysis in the herbage, which has led to increases in N use efficiency and polyunsaturated fatty acid (PUFA) content of milk when offered to dairy cows. In damaged plant cells, PPO is activated and binds protein through the formation of protein-bound phenols. This study investigated a) whether freshly cut red clover could increase N use efficiency and milk PUFA concentrations in dairy cows or whether PPO enzymes require prior activation before feeding to elicit a response, and b) apparent whole-tract amino acid digestibility to help determine the effect of PPO on amino acid utilization. Six multiparous Holstein×Friesian dairy cows in mid-lactation were allocated at random to 1 of 3 dietary treatments in a 3×3 Latin square: a control treatment of grass (low PPO, G); red clover (high PPO, RC), and conditioned red clover (high fully activated PPO, CRC). The CRC herbage was cut and chopped in the field and then transported with the G and RC herbages to the animal house. Each period consisted of a 2-wk adaptation to diet and a week of measuring dietary effects (N balance and milk collection). The PPO activity was greatest in the RC treatment as fed, whereas activation of latent PPO enzyme and protein-bound phenol levels were greatest in the CRC diet. Dry matter and total fatty acid intakes were comparable across treatments (18.8kg/d and 550g/d, respectively). Milk yields and total fatty acid content were similar across treatments (32.6kg/d and 34.8mg/mL, respectively). Cows offered either RC or CRC had greater levels of protein, C18 PUFA and total long-chain PUFA in their milk than animals offered grass with no difference between RC and CRC. Nitrogen intakes, and output in milk, urine, and feces were greater in cows offered the 2 red clover treatments than G, with no difference between RC and CRC. However, there were no differences in N use efficiency among diets as measured by the proportion of feed N converted into milk N, possibly as the result of the excessive supply of N with the red clover diets. Amino acid apparent whole-tract digestibilities were greater when on RC than G diets and intermediate when on CRC for all amino acids, with the exception of Met, which was reduced in cows on both red clover diets compared with G. It is proposed that the PPO trait could show more benefit to ruminants if red clover was fed in combination with lesser N-containing forages or if red clover was bred to contain less N.