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The nonprecious electrocatalyst with high electrocatalytic activity and low cost is urgently desired to fulfill the demands of efficient rechargeable Zn-air batteries (ZABs). Herein, the heteroatom functional carbon-based nanocage Zn-NC is prepared through pyrolysis of ZIF-8. In this process, the organic ligands are converted into partially graphitized carbon with simultaneously reduction of Zn2+ to metal Zn, thus the nitrogen doping porous structure and homogeneous distributed zinc endow the Zn-NC more exposed active surface. Further co-heat treatment of carbonized ZIF-8 precursor with dicyandiamide and glucose results in the formation of Zn, N atomic-level modified carbon material Zn-NC/GD. The dual-shell carbon nanocage structure produces obvious interface properties, which can well bind the escaping metallic zinc to form Zn–N–C bond structure. Benefiting from double carbon modification, interfacial properties, and bonding structure, the Zn-NC/GD performs outstanding half-wave potential (0.86 V) and a lower Tafel slope (97.8 mV dec−1). Consequently, as an air electrode, the Zn-NC/GD also exhibits high open circuit voltage (1.531 V), remarkable power density (326.1 mW cm−2), and excellent specific capacity (824.7 mAh gzn−1). The critical findings demonstrate promising potential to obtain an efficient atomic-level modification of carbon-based electrocatalyst for ZABs.

Dietary supplementation with 0.4 or 0.8% l-arginine (Arg) to gilts between days 14 and 25 of gestation enhances embryonic survival and vascular development in placentae; however, the underlying mechanisms are largely unknown. This study tested the hypothesis that Arg supplementation stimulated placental expression of mRNAs and proteins that enhance angiogenesis, including endothelial nitric oxide synthase (eNOS), vascular endothelial growth factor (VEGF), placental growth factor (PGF), GTP cyclohydrolase-I (GTP-CH1), ornithine decarboxylase (ODC1), and vascular endothelial growth factor receptors 1 and 2 (VEGFR1 and VEGFR2). Beginning on the day of breeding, gilts were fed daily 2 kg of a corn–soybean meal-based diet supplemented with 0.0 (control), 0.4, or 0.8% Arg. On day 25 of gestation, gilts were hysterectomized to obtain uteri and conceptuses for histochemical and biochemical analyses. eNOS and VEGFR1 proteins were localized to endothelial cells of maternal uterine blood vessels and to the uterine luminal epithelium, respectively. Compared with the control, dietary supplementation with 0.4 or 0.8% Arg increased (P

Electronic storage devices assembled from traditional hydrogel electrolytes (HEs) lose most of their capacity due to low-temperature effects and fail at high temperatures due to the water evaporati...

As the first line of defence against pathogens and endotoxins crossing the intestine-blood barrier, the intestinal epithelial barrier plays a determinant role in pigs' health and growth. 4-Phenylbutyric acid (4-PBA), an aromatic fatty acid, was reported to benefit homeostasis of endoplasmic reticulum and protein synthesis. However, whether 4-PBA affects intestinal epithelial barrier function in pigs is unknown. This study aimed to explore the effects of 4-PBA on the intestinal barrier function, using in vitro models of well-differentiated intestinal porcine epithelial cell (IPEC-J2) monolayers in the transwell plates. Cell monolayers with or without 4-PBA (1.0 mmol/L) treatment were challenged with physical scratch, deoxynivalenol (DON, 2.0 μg/mL, 48 h), and lipopolysaccharide (LPS, 5.0 μg/mL, 48 h), respectively. Transepithelial electrical resistance (TEER) and fluorescein isothiocyanate-dextran (FD-4) permeability were measured to indicate barrier integrity and permeability. Real-time PCR and Western blot were conducted to determine relative gene and protein expressions of tight junction proteins. As expected, physical scratch, DON, and LPS challenges decreased TEER and increased FD-4 permeability. 4-PBA treatment accelerated cell mitigation and rehabilitation of the physical scratch-damaged intestinal epithelial barrier but did not alleviate DON or LPS induced barrier damage. However, once 48-h DON and LPS challenges were removed, rehabilitation of the epithelial barrier function of IPEC-J2 monolayer was accelerated by the 4-PBA treatment. Also, the relative gene and protein expressions of zonula occludens-1 (ZO-1), occludin, and claudin-1 were further upregulated by the 4-PBA treatment during the barrier rehabilitation. Taken together, 4-PBA accelerated the IPEC-J2 cell monolayer barrier recovering from physical scratch, DON-, and LPS-induced damage, via enhancing cell mitigation and expressions of tight junction proteins.

Due to multiple compensating mechanisms, the serum bicarbonate concentration is a relatively insensitive marker of acid-base status; especially in chronic kidney disease (CKD). This is a major drawback that impairs the ability to diagnose acid excess or monitor alkali therapy. We postulated that it is more logical to measure the compensatory defense mechanism(s) rather than the defended parameter, which remains normal if the compensation is successful. Therefore, a retrospective cross-sectional study was performed in 1733 stone formers along with a prospective cross-sectional study of 22 individuals with normal kidney function and 50 patients in different stages of CKD. While serum bicarbonate was flat and did not fall below the reference range until near CKD stage 5, citrate excretion (24-hour urinary citrate excretion rate; urinary citrate-to-creatinine ratio, in the retrospective analysis, and spot urinary citrate-to-creatinine ratio in the prospective study) progressively and significantly declined starting from CKD stage 2. Following an acute acid load in 25 participants with a wide range of estimated glomerular filtration rates, the urinary citrate-to-creatinine ratio inversely and significantly associated with acid accumulation, whereas serum bicarbonate did not. We compared changes in serum bicarbonate and urinary citrate-to-creatinine ratio in response to alkali therapy in patients with CKD stage 3 or 4 started on potassium citrate in our kidney stone database. With alkali therapy, there was no change in serum bicarbonate, but the urinary citrate-to-creatinine ratio rose consistently in all patients adherent to potassium citrate therapy. Thus, the urinary citrate-to-creatinine ratio (the defense mechanism) is a potential easily implementable, pragmatic, and a superior parameter to serum bicarbonate (the defended entity) to assess acid-base status, and monitor alkali therapy. Additional studies are needed before a clinical test can be devised.

Soybean meal is rich in soybean isoflavones, which exhibit antioxidant, anti-inflammatory, antiviral and anticancer functions in humans and animals. This study was conducted to investigate the effects of soybean isoflavones on the growth performance, intestinal morphology and antioxidative properties in pigs. A total of 72 weaned piglets (7.45 ± 0.13 kg; 36 males and 36 females) were allocated into three treatments and fed corn-soybean meal (C-SBM), corn-soy protein concentrate (C-SPC) or C-SPC supplemented with equal levels of the isoflavones found in the C-SBM diet (C-SPC + ISF) for a 72-day trial. Each treatment had six replicates and four piglets per replicate, half male and half female. On day 42, one male pig from each replicate was selected and euthanized to collect intestinal samples. The results showed that compared to pigs fed the C-SPC diet, pigs fed the C-SBM and C-SPC + ISF diets had higher BW on day 72 (P < 0.05); pigs fed the C-SBM diet had significantly higher average daily gain (ADG) during days 14 to 28 (P < 0.05), with C-SPC + ISF being intermediate; pigs fed the C-SBM diet tended to have higher ADG during days 42 to 72 (P = 0.063), while pigs fed the C-SPC + ISF diet had significantly higher ADG during days 42 to 72 (P < 0.05). Moreover, compared to pigs fed the C-SPC diet, pigs fed the C-SBM diet tended to have greater villus height (P = 0.092), while pigs fed the C-SPC + ISF diet had significantly greater villus height (P < 0.05); pigs fed the C-SBM and C-SPC + ISF diets had significantly increased villus height-to-crypt depth ratio (P < 0.05). Compared with the C-SPC diet, dietary C-SPC + ISF tended to increase plasma superoxide dismutase activity on days 28 (P = 0.085) and 42 (P = 0.075) and reduce plasma malondialdehyde (MDA) content on day 42 (P = 0.089), as well as significantly decreased jejunal mucosa MDA content on day 42 (P < 0.05). However, no significant difference in the expression of tight junction genes among the three groups was found (P > 0.05). In conclusion, our results suggest that a long-term exposure to soybean isoflavones enhances the growth performance, protects the intestinal morphology and improves the antioxidative properties in pigs.

Background: Persistence of acute kidney disease (AKD) after an episode of acute kidney injury (AKI) is associated with adverse outcomes. Multiple factors contribute to AKD after AKI, but the role of angiotensin-converting enzyme inhibitors/angiotensin II receptor blockers (ACEI/ARB) remains controversial. We examined if acute exposure to an ACEI/ARB associates with persistent AKD in survivors of AKI. Methods: Multicenter prospective cohort study of patients whose hospitalization was complicated by AKI and who attended specialized AKI follow-up clinics between 2013 and 2018. Acute exposure was defined as ACEI/ARB exposure for ≥48 h before or during the AKI episode. The primary outcome was AKD (serum creatinine ≥1.5 times above pre-AKI baseline) at the first clinic visit. We used multivariable logistic regression to adjust for potential confounders. Results: We included 345 survivors of AKI, 112 with persistent AKD at the first outpatient visit. Among 163 patients who were prescribed an ACEI/ARB before hospitalization, only 23% were discharged on an ACEI/ARB. There was no difference in the rate of AKD in patients discharged versus not discharged on an ACEI/ARB (12.5 vs. 15.0%, p = 0.530). Of the patients with AKD, 22 (19.6%) patients had acute ACEI/ARB exposure during the hospitalization. In fully adjusted models, acute exposure to an ACEI/ARB was not associated with AKD at the time of first clinic visit (median [interquartile range] 33 [18–54] days from hospital discharge). Conclusion: Acute exposure to an ACEI/ARB before or during an episode of AKI was not associated with persistent AKD at the time of first clinic visit suggesting that the receipt of such agents does not impede kidney recovery following AKI. Contrary to prevailing recommendations and current practice, the continued administration of an ACEI/ARB during an episode of AKI or initiation of these agents prior to discharge may be safe.

Introduction Our aim was to evaluate whether mechanical forces applied to the placenta of pigs correlate with morphological changes that coordinate the development of placental folds. Methods We examined changes in the length of placental folds, expression of mechanotransduction-implicated molecules in placental tissues, changes in the size of subepithelial blood vessels within the endometrium, and effects of in vivo supplementation with arginine on fold development. Results We observed that: 1) the length of folds increased 2) osteopontin, talin and focal adhesion kinase co-localized into aggregates at the maternal placental (uterine)-fetal placental interface; 3) filamin, actin related protein 2, and F-actin were enriched in the tops of maternal placental folds extending into fetal placental tissue; 4) maternal stromal fibroblasts acquired alpha smooth muscle actin; 5) endometrial blood vessels increased in size; and 6) supplementation with arginine increased fold length. Conclusion Results indicate that lengthening of folds associates with polymerization of actin that coincides with FA assembly, endometrial fibroblasts differentiate into myofibroblasts, and dilation of subepithelial blood vessels correlates with development of folds that is enhanced by arginine. We propose that dilation of subepithelial endometrial blood vessels delivers increased blood flow that pushes upward on the interface between the uterine luminal epithelium (LE) and the placental chorionic epithelium (CE), protrusive forces from growing uterine blood vessels trigger focal adhesion assembly and actin polymerization between the LE and CE, and endometrial fibroblasts differentiate into contractile myofibroblasts that pull connective tissue downward and inward to sculpt folds at the maternal placental-fetal placental interface.

Bismuth has a great potential as an anode for PIBs, due to its layered structure with a large interlayer spacing, long mean free path, high volumetric capacity (3800 mA h L−1) and environmental friendliness. Bismuthene, a 2D-layered structure, can be exfoliated from bismuth by mechanical or chemical methods. The unique structure of bismuthene is beneficial to the diffusion of potassium, penetration of the electrolyte, and buffering of the volume change along the c-axis, which will boost the anode performance of PIBs. An ultrasonication-assisted electrochemical exfoliation method was proposed in this study to prepare ultra-thin few layered bismuthene nanosheets (FBNs) simply and quickly. The as-prepared FBNs were employed in KIBs as an anode and delivered highly stable capacities of 423, 356, 275 and 227 mA h g−1 at the current densities of 2.5, 5, 10, and 15 A g−1, respectively. There was no obvious decay over 2500 cycles with a capacity of over 200 mA h g−1 at 20 A g−1, realizing excellent rate-capability and long-term cycling stability in KIBs. Furthermore, the mechanism of the excellent electrochemical performances was elucidated via in-depth characterization and theoretical calculations. This work provides a new strategy to prepare scalable ultra-thin nanosheets, which act as a promising candidate for the anode in energy storage systems.

Simple Summary Gallic acid (GA) has been demonstrated to have antioxidant, antimicrobial, anti-inflammatory, and health-promoting properties. In pigs, GA supplementation has been shown to decrease di-arrhea incidence of weaned piglets and improve their intestinal integrity. The present experiment was conducted to test the hypothesis that growth performance and diarrhea after weaning could be improved by supplementing the diet with 400 mg/kg GA to weaned piglets, especially for low weaning weight piglets. Abstract This study evaluated the effects of dietary gallic acid (GA) on growth performance, diarrhea incidence and plasma antioxidant status of weaned piglets regardless of whether weaning weight was high or low. A total of 120 weaned piglets were randomly allocated to four treatments in a 42-day experiment with a 2 × 2 factorial treatment arrangement comparing different weaning weights (high weight (HW) or low weight (LW), 8.49 ± 0.18 kg vs. 5.45 ± 0.13 kg) and dietary treatment (without supplementation (CT) or with supplementation of 400 mg/kg of GA). The results showed that HW piglets exhibited better growth performance and plasma antioxidant capacity. Piglets supplemented with GA had higher body weight (BW) on day 42 and average daily gain (ADG) from day 0 to 42 compared to the control piglets, which is mainly attributed to the specific improvement on BW and ADG of LW piglets by the supplementation of GA. The decreased values of diarrhea incidence were seen in piglets fed GA, more particularly in LW piglets. In addition, dietary GA numerically reduced malondialdehyde (MDA) content in plasma of LW piglets. In conclusion, our study suggests that dietary GA may especially improve the growth and health in LW weaned piglets.

This study was conducted to investigate the effect of fermented Radix puerariae residue (FRPR) on reproductive performance, apparent total tract digestibility (ATTD) of nutrients, and fecal short-chain fatty acid (SCFA) contents of sows. A total of 36 landrace × large white multiparous sows were randomly arranged into three treatments, representing supplementation with 0, 2, and 4% FRPR to a corn-soybean meal and wheat bran-based diet during the whole gestation period. The results showed that dietary FRPR had no effects on litter size and the number of total alive piglets (P > 0.05), and that the number of weaned piglets and weaning weight of litter were increased in sows with 4% FRPR treatment compared with control treatment (P < 0.05). Dietary 4% FRPR significantly decreased constipation rate, improved the ATTD of dry matter and organics, and fecal contents of acetate, propionate, and total SCFAs (P < 0.05). In the offspring piglets, serum concentrations of total protein, alkaline phosphatase, IgG, IL-10, and TGF-β were increased, but blood urea nitrogen content was decreased with 4% FRPR treatment (P < 0.05). There were no significant differences in all determined indexes except for fecal acetic acid and total SCFAs between control and 2% FRPR treatment (P > 0.05). These findings indicated that FRPR used in the diets of sows showed positive effects on fecal characteristics, utilization of nutrients, and reproductive performance. Maternal supplementation with 4% FRPR is recommended for improving immune responses, weaning litter size, and litter weight of offspring piglets, which provide useful information for the application of residues of R. puerariae.

Hypophosphatemia is a common and dangerous complication of acute liver failure (ALF) of various etiologies. While various mechanisms for ALF-associated hypophosphatemia have been proposed including high phosphate uptake into regenerating hepatocytes, acetaminophen (APAP)-associated hypophosphatemia was linked to renal phosphate wasting, and APAP-induced renal tubular injury was proposed as underlying mechanism. We studied 30 normophosphatemic and 46 hypophosphatemic (serum phosphate < 2.5 mg/dL) patients from the Acute Liver Failure Study Group registry with APAP- or non-APAP-induced ALF. Since kidney injury affects phosphate excretion, patients with elevated serum creatinine (>1.2 mg/dL) were excluded. Maximal amount of renal tubular phosphate reabsorption per filtered volume (TmP/GFR) was calculated from simultaneous serum and urine phosphate and creatinine levels to assess renal phosphate handling. Instead of enhanced renal phosphate reabsorption as would be expected during hypophosphatemia of non-renal causes, serum phosphate was positively correlated with TmP/GFR in both APAP- and non-APAP-induced ALF patients (R(2) = 0.66 and 0.46, respectively; both p < 0.0001), indicating renal phosphate wasting. Surprisingly, there was no evidence of kidney damage based on urinary markers including neutrophil gelatinase-associated lipocalin (NGAL), and cystatin C even in the APAP group. Additionally, there was no evidence that the known serum phosphatonins parathyroid hormone (PTH), fibroblast growth factor 23 (FGF23), and α-Klotho contribute to the observed hypophosphatemia. We conclude that the observed hypophosphatemia with renal phosphate wasting in both APAP- and non-APAP-mediated ALF is likely the result of renal tubular phosphate leak from yet-to-be identified factor(s) with no evidence for proximal tubular damage or contribution of known phosphatonins.

Two-dimensional MXene Ti3C2T x nanosheets with peroxide decoration (p-Ti3C2T x ) are synthesized by a sonication-assisted MILD etching method. The obtained MXenes can generate hydroxyl radical species and act as an initiator for free-radical polymerization of a series of acrylic monomers without the use of light illumination or co-initiators. The monomers analyzed include acrylamide, N-isopropylacrylamide (NIPAM), N,N-dimethylacrylamide, methyl methacrylate, and hydroxyethyl methacrylate. By simply mixing N-isopropylacrylamide monomers and p-Ti3C2T x nanosheets under deoxygenated conditions, PNIPAM-based nanocomposite hydrogels are synthesized using a high concentration of the monomer. The nanocomposite hydrogels have a photothermal conversion efficiency of 34.7% and photothermal stability superior to that of pristine Ti3C2T x . Taking advantage of the thermal responsive behavior of PNIPAM, the nanocomposite hydrogels are successfully exploited as remotely near-infrared light controlled "smart" windows, fluidic valves and photodetectors.

Gallic acid is a phenolic compound that exhibits antibacterial, antioxidative and anti-inflammatory functions. In a previous study, we found that dietary supplementation with gallic acid decreased incidence of diarrhoea and protected intestinal integrity in weaning piglets. However, the underlying mechanism remains unclear. Here, a pig intestinal epithelial cell line (IPEC-J2) was used as an in vitro model to explore the antioxidant and anti-inflammatory capacity of gallic acid. IPEC-J2 cells were stimulated with hydrogen peroxide (H2 O2 ) and lipopolysaccharide (LPS) to establish oxidative and inflammatory models, respectively. Results showed that H2 O2 significantly decreased catalase (CAT) secretion and CAT mRNA abundance in the cells (p < 0.05), while pretreatment with gallic acid did not prevent the decrease in CAT expression induced by H2 O2 . However, gallic acid pretreatment mitigated the increased expression of the tumour necrosis factor-α and interleukin-8 genes caused by LPS in IPEC-J2 cells (p < 0.05). In addition, pretreatment with gallic acid significantly suppressed phosphorylation of NF-κB and IκBα in LPS-stimulated IPEC-J2 cells. Moreover, LPS stimulation decreased the protein abundance of zona occludens 1 (ZO-1) and occludin, while pretreatment with gallic acid preserved expression level of tight junction proteins ZO-1 and occludin in LPS-stimulated IPEC-J2 cells (p < 0.05). In conclusion, gallic acid may mitigate LPS-induced inflammatory responses by inhibiting the NF-κB signalling pathway, exerting positive effects on the barrier function of IPEC-J2 cells.

Nephrolithiasis is associated with an increased risk of chronic kidney disease, and its incidence varies with age. However, little is known on the combined impact of aging and declining renal function on urinary risk factors for calcium oxalate stone formation. A retrospective analysis was performed on 24-hour urine collections from 993 calcium oxalate stone-forming patients. We first tested for interactions between age and creatinine clearance on various urinary determinants of calcium oxalate nephrolithiasis, and then examined their separate and combined effects in univariable and multivariable analyses adjusting for demographic and biochemical covariates. We identified significant interactions between age and creatinine clearance in predicting 24-hour urine pH, calcium, and citrate. In view of the small number of stone formers with low creatinine clearance, we limited further regression analyses to patients with creatinine clearance ≥60 mL/min. In multivariable analyses, urine citrate, oxalate, and total volume were positively correlated with age, whereas urine pH, citrate, calcium, oxalate, total volume, and RSR of calcium oxalate all significantly decreased with lower creatinine clearance. A decrease in creatinine clearance from 120 to 60 ml/min was associated with clinically significant decreases in the daily excretion rate of citrate (by 188 mg/day), calcium (by 33 mg/day), and oxalate (by 4 mg/day), and in RSR calcium oxalate (by 1.84). Age and creatinine clearance are significant and independent predictors of several urinary determinants of calcium oxalate nephrolithiasis. The impacts of aging and declining renal function should be considered during the management of calcium oxalate stone-forming patients.

Background: In previous study, we found that soybean isoflavones in soybean meal play important roles in improving growth performance and antioxidant properties in pigs. However, it is still not known whether long-term supplementation with daidzein, an active molecule deglycosylated from daidzin, in a corn-soybean meal diet can enhance growth performance in pigs. Thus, in the present study, an animal trial was carried out to investigate the effects of dietary supplementation with daidzein on the growth performance and antioxidant capacity of pigs. Porcine intestinal epithelial cells (IPEC-J2) were also used as an in vitro model to explore the underlying antioxidant mechanisms of daidzein. Weanling pigs were fed a diet supplemented with 0, 25, 50, and 100 mg/kg daidzein, and IPEC-J2 cells were treated with 0.6 mM hydrogen peroxide (H2O2) in the presence or absence of 40 μM daidzein. Results: Adding 50 mg/kg daidzein to the diet significantly improved body weight on day 72, average daily gain during days 0-72 and plasma superoxide dismutase activity on day 42 (P < 0.05). Treatment with 0.6 mM H2O2 for 1 h significantly decreased cell viability and catalase (CAT) activities and increased intracellular reactive oxygen species (ROS) levels and malondialdehyde (MDA) content (P < 0.05), while pretreatment with 40 μM daidzein prevented the decrease in cell viability and CAT activities and the increase in intracellular ROS levels and MDA content caused by H2O2 (P < 0.05). In addition, H2O2 stimulation significantly suppressed the expression of nuclear factor erythoid-2-related factor 2 (Nrf2), CAT, occludin and zonula occludens-1 (ZO-1), while pretreatment with daidzein preserved the expression of Nrf2, CAT, occludin and ZO-1 in H2O2-stimulated IPEC-J2 cells (P < 0.05).Conclusions: Long-term dietary supplementation with 50 mg/kg daidzein improved growth performance and antioxidative properties in pigs. Daidzein exerted protective effects against H2O2-induced oxidative stress in IPEC-J2 cells, and the underlying mechanism may be related to the activation of the Kelch-like ECH-associated protein 1-Nrf2/antioxidant response element signaling pathway.

Background Polyamines are essential for cell growth and beneficial for intestinal maturation. To evaluate the effects of putrescine on alleviating intestinal atrophy and underlying molecular mechanisms, both in vivo feeding trial and in vitro cell culture were conducted. Weanling pigs were fed a diet supplemented with 0, 0.1%, 0.2% or 0.3% putrescine dihydrochloride, whereas porcine intestinal epithelial cells (IPEC-J2) were challenged with lipopolysaccharide (LPS) in the presence of 200 μmol/L putrescine. Results Dietary supplementation with 0.2% putrescine dihydrochloride decreased the incidence of diarrhea with an improvement in intestinal integrity. Inhibition of ornithine decarboxylase activity decreased the proliferation and migration of IPEC-J2 cells, and this effect was alleviated by the supplementation with putrescine. The phosphorylation of extracellular signal regulated kinase and focal adhesion kinase was enhanced by putrescine. LPS increased the expression of inflammatory cytokines [tumor necrosis factor α (TNF-α), interleukin 6 (IL-6) and IL-8], and inhibited cell proliferation and migration in IPEC-J2 cells. Adding exogenous putrescine suppressed the expression of TNF-α, IL-6 and IL-8, and recovered cell migration and proliferation in LPS-treated IPEC-J2 cells. Dietary putrescine supplementation also reduced the mRNA levels of TNF-α, IL-6 and IL-8 and their upstream regulator nuclear receptor kappa B p65 subunit in the jejunal mucosa of piglets. Conclusions Dietary supplementation with putrescine mitigated mucosal atrophy in weanling piglets through improving anti-inflammatory function and suppressing inflammatory response. Our results have important implications for nutritional management of intestinal integrity and health in weanling piglets and other neonates.

Peroxide-decorated Ti3C2Tx plays a role as an initiator for free-radical polymerization and leads to new opportunities for intelligent optoelectronic devices., Two-dimensional MXene Ti3C2Tx nanosheets with peroxide decoration (p-Ti3C2Tx) are synthesized by a sonication-assisted MILD etching method. The obtained MXenes can generate hydroxyl radical species and act as an initiator for free-radical polymerization of a series of acrylic monomers without the use of light illumination or co-initiators. The monomers analyzed include acrylamide, N-isopropylacrylamide (NIPAM), N,N-dimethylacrylamide, methyl methacrylate, and hydroxyethyl methacrylate. By simply mixing N-isopropylacrylamide monomers and p-Ti3C2Tx nanosheets under deoxygenated conditions, PNIPAM-based nanocomposite hydrogels are synthesized using a high concentration of the monomer. The nanocomposite hydrogels have a photothermal conversion efficiency of 34.7% and photothermal stability superior to that of pristine Ti3C2Tx. Taking advantage of the thermal responsive behavior of PNIPAM, the nanocomposite hydrogels are successfully exploited as remotely near-infrared light controlled “smart” windows, fluidic valves and photodetectors.

AIM Acute kidney injury (AKI) is a frequent complication of sepsis, a pro-inflammatory state that alters tubular handling of filtered albumin. We hypothesized that dipstick albuminuria (DA) is associated with a lower rate of AKI recovery in septic patients. METHODS This was a single-centre, retrospective cohort study of adults with sepsis-associated AKI in an urban academic intensive care unit (ICU). Patients with unknown baseline serum creatinine (SCr), absent urinalysis, and those with estimated glomerular filtration rate (eGFR)

Glutathione (GSH) forms conjugates with polyamines in prokaryotes and eukaryotes. There is also evidence suggesting cross-talk between GSH and polyamines to regulate cellular homeostasis and function, particularly under the conditions of oxidative stress. Because of its versatile roles in cell metabolism and function, a number of high performance liquid chromatography (HPLC) methods have been developed for glutathione analysis. Here, we describe our rapid and sensitive method for the analysis of GSH and the oxidized form of glutathione (GS-SG) in animal tissues and cells by HPLC involving pre-column derivatization with o-phthalaldehyde (OPA). OPA reacts very rapidly (within 1 min) with S-carboxymethyl-glutathione at room temperatures (e.g., 20-25 °C) in an autosampler, and their derivatives are immediately injected into the HPLC column without any need for extraction. This method requires two simple steps (a total of 15 min) before samples are loaded into the autosampler: (a) the conversion of GS-SG into GSH by 2-mercaptoethanol; and (b) the oxidation of GSH by iodoacetic acid to yield S-carboxymethyl-glutathione. The autosampler is programmed to mix S-carboxymethyl-glutathione with OPA for 1 min to generate a highly fluorescent derivative for HPLC separation and detection (excitation wavelength 340 nm and emission wavelength 450 nm). The detection limit for GSH and GS-SG is 15 pmol/ml or 375 fmol/injection. The total time for chromatographic separation (including column regeneration) is 16 min for each sample. Our routine HPLC technique is applicable for analyses of cysteine and cystine, as well as polyamines and GSH-polyamine conjugates in biological samples.

Supplemental Digital Content is available in the text., Objectives: Klotho and fibroblast growth factor-23 were recently postulated as candidate biomarkers and/or therapeutic targets in acute kidney injury. We examined whether urine Klotho and serum intact fibroblast growth factor-23 levels were differentially and independently associated with major adverse kidney events in critically ill patients with and without acute kidney injury. Design: Single-center, prospective, case-control study. Setting: ICU in a tertiary medical center. Patients: Fifty-four acute kidney injury patients and 52 controls without acute kidney injury admitted to the ICU. Interventions: None. Measurements and Main Results: Acute kidney injury was defined by Kidney Disease: Improving Global Outcomes criteria and included only acute kidney injury stage greater than or equal to 2. Controls were matched by age, gender, and baseline estimated glomerular filtration rate. Paired serum and urine samples were obtained 24–48 hours after acute kidney injury diagnosis (cases) or ICU admission (controls). The primary outcome was 90-day major adverse kidney events, which was the composite of all-cause death, dependence on renal replacement therapy, or a 50% or higher decrease in estimated glomerular filtration rate from baseline. Major adverse kidney events, which was the composite of all-cause death, dependence on renal replacement therapy, or a 50% or higher decrease in estimated glomerular filtration rate from baseline. Major adverse kidney events developed in 44 patients (41.5%). Patients in whom major adverse kidney events developed had more comorbidity, higher acuity of illness scores, and more prevalent acute kidney injury. Levels of urine Klotho adjusted by creatinine were lower, and serum intact fibroblast growth factor-23 levels were higher in acute kidney injury patients versus ICU controls. In adjusted models, the highest versus lowest tertile of urine Klotho/creatinine was independently associated with an overall 95% lower risk of major adverse kidney events (81% lower risk in patients with acute kidney injury). The highest versus lowest tertile of serum intact fibroblast growth factor-23 was associated with more than 300% higher risk of major adverse kidney events. Conclusions: Urine Klotho/creatinine levels were significantly lower and serum intact fibroblast growth factor-23 levels were significantly higher in critically ill patients with acute kidney injury versus matched controls without acute kidney injury. When measured in the first 48 hours of ICU admission or acute kidney injury diagnosis, urine Klotho/creatinine independently associated with major adverse kidney events, particularly in patients with acute kidney injury. These results show promise for testing these biomarkers—individually or in combination—as part of novel risk prediction models of renal outcomes in the ICU.

Male Zucker diabetic fatty (ZDF) rats were used to study effects of oral administration of interferon tau (IFNT) in reducing obesity. Eighteen ZDF rats (28 days of age) were assigned randomly to receive 0, 4 or 8 μg IFNT/kg body weight (BW) per day (n=6/group) for 8 weeks. Water consumption was measured every two days. Food intake and BW were recorded weekly. Energy expenditure in 4-, 6-, 8-, and 10-week-old rats was determined using indirect calorimetry. Starting at 7 weeks of age, urinary glucose and ketone bodies were tested daily. Rates of glucose and oleate oxidation in liver, brown adipose tissue, and abdominal adipose tissue, leucine catabolism in skeletal muscle, and lipolysis in white and brown adipose tissues were greater for rats treated with 8 μg IFNT/kg BW/day in comparison with control rats. Treatment with 8 μg IFNT/kg BW/day increased heat production, reduced BW gain and adiposity, ameliorated fatty liver syndrome, delayed the onset of diabetes, and decreased concentrations of glucose, free fatty acids, triacylglycerol, cholesterol, and branched-chain amino acids in plasma, compared to control rats. Oral administration of 8 μg IFNT/kg BW/day ameliorated oxidative stress in skeletal muscle, liver and adipose tissue, as indicated by decreased ratios of oxidized glutathione to reduced glutathione and increased concentrations of the antioxidant tetrahydrobiopterin. These results indicate that IFNT stimulates oxidation of energy substrates and reduces obesity in ZDF rats and may have broad important implications for preventing and treating obesity-related diseases in mammals.

The gas molecule nitric oxide (NO) can cooperate with ethylene to tightly modulate plant growth and stress responses. One of the mechanism of their crosstalk is that NO is able to activate ethylene biosynthesis, possibly through post-translational modification of key enzymes such as ACC synthase and oxidase by S-nitrosylation. In this paper, we focus on the crosstalk of NO with ethylene signaling transduction transcription factor EIN3 (Ethylene Insensitive 3) and downstream gene expression in alleviating germination inhibition and growth damage induced by high salt. The Arabidopsis lines affected in ethylene signaling (ein3eil1) and NO biosynthesis (nia1nia2) were employed to compare with the wild-type Col-0 and overexpressing line EIN3ox. Firstly, the obviously inhibited germination, greater ratio of bleached leaves and enhanced electrolyte leakage were found in ein3eil1 and nia1nia2 lines than in Col-0 plants upon high salinity. However, the line EIN3ox obtained a notably elevated ability to germinate and improved seedling resistance. The experiment with SNP alone or plus high salt mostly enhanced the expression of EIN3 transcripts, compared with ACO4 and ACS2. The western blot and transcript analysis found that high-salt-induced EIN3 stabilization and EIN3 transcripts were largely attenuated in the NO biogenesis mutant nia1nia2 plants than in Col-0 ones. This observation was confirmed by simulation experiments with NO scavenger cPTIO to block NO emission. Taken together, our study provides insights that NO promotes seed germination and seedlings growth under salinity may depend on EIN3 protein.

We proposed that the combination of conjugated linoleic acid (CLA) and arginine would decrease adiposity by depressing lipid synthesis in liver and adipose tissues of growing pigs. Pigs were allotted to treatments in a 2 × 2 factorial design with two lipids (CLA or canola oil) and two amino acids [L-arginine or L-alanine (isonitrogenous control)]; supplements were provided from 80 to 110 kg body weight (approximately 4 weeks). Treatment groups (n = 4) were: control (2.05% L-alanine plus 1% canola oil); CLA (2.05% L-alanine plus 1% CLA); arginine (1.0% L-arginine plus 1.0% canola oil); arginine plus CLA (1.0% arginine plus 1.0% CLA). Arginine increased backfat thickness (P = 0.07) in the absence or presence of CLA, and arginine supplementation increased subcutaneous and retroperitoneal adipocyte volume, especially in combination with dietary CLA (interaction P = 0.001). Arginine increased palmitate incorporation into total lipids by over 60% in liver (P = 0.07). Dietary CLA increased palmitate incorporation into lipids in longissimus muscle by over 100% (P = 0.01), and CLA increased longissimus muscle lipid by nearly 20%. CLA increased glucose oxidation to CO(2) by over 80% in retroperitoneal and subcutaneous adipose tissues (P = 0.04), and doubled palmitate oxidation to CO(2) in intestinal duodenal mucosal cells (P = 0.07). Arginine supplementation decreased muscle pH at 45 min postmortem (P = 0.001), indicating elevated early postmortem glycolysis, and CLA and arginine independently increased PGC-1α gene expression in longissimus muscle. CLA but not arginine depressed mTOR gene expression in intestinal duodenal mucosal cells. CLA decreased serum insulin by 50% (P = 0.02) but increased serum triacylglycerols by over 40%. CLA supplementation increased (P ≤ 0.01) total saturated fatty acids in liver and adipose tissue. In conclusion, neither CLA nor arginine depressed tissue lipid synthesis in growing/finishing pigs, and in fact dietary CLA promoted elevated intramuscular lipid and arginine increased carcass adiposity.

This study was to test the hypothesis that altered IGF2 system in the placental labyrinth zone (LZ) impairs feto-placental growth in response to maternal protein restriction. Rats were fed a 20% protein diet and an isocaloric 6 % protein diet (LP) from day 1 to days 14, 18, or 21 of pregnancy. The effects of diet, gender of placenta and fetus, and day of pregnancy on placental weight, fetal weight, and expression of the IGF2 axis in the placental LZ and amino acids in maternal plasma were analyzed. Growth restriction occurred in both female and male fetuses by LP, coincident with impaired LZ growth and efficiency. The expression of Igf2, Igf2P0, Igf1r, Igf2r, Insr, Igfbp1, and Igfbp2 in placental LZ were affected by diet, gender and/or day of pregnancy. Concentrations of total essential amino acids and total nonessential amino acids were reduced and increased, respectively, in maternal plasma of LP-fed rats. These results indicate that adaptation of the IGF2 system in rat LZ occurs in a sex- and time-dependent manner in response to maternal protein restriction; however, these adaptations cannot prevent the growth restriction of both male and female fetuses during late pregnancy.

Pigs suffer up to 50% embryonic and fetal loss during gestation and exhibit the most severe naturally occurring intrauterine growth retardation among livestock species. Placental insufficiency is a major factor contributing to suboptimal reproductive performance and reduced birth weights of pigs. Enhancement of placental growth and function through nutritional management offers an effective solution to improving embryonic and fetal survival and growth. We discovered an unusual abundance of the arginine family of AA in porcine allantoic fluid (a reservoir of nutrients) during early gestation, when placental growth is most rapid. Arginine is metabolized to ornithine, proline, and nitric oxide, and these compounds possess a plethora of physiological functions. Nitric oxide is a vasodilator and angiogenic factor, whereas both ornithine and proline are substrates for placental synthesis of polyamines, which are key regulators of protein synthesis and angiogenesis. Additionally, arginine, leucine, glutamine, and proline activate the mammalian target of rapamycin cell-signaling pathway to enhance protein synthesis and cell proliferation in placentae. To translate basic research on AA biochemistry and nutrition into application, dietary supplementation with 0.83% l-arginine to gilts on d 14 to 28 or d 30 to 114 of gestation increased the number and litter birth weight of live-born piglets. In addition, supplementing the gestation diet with 0.4% l-arginine plus 0.6% l-glutamine enhanced the efficiency of nutrient utilization, reduced variation in piglet birth weight, and increased litter birth weight. By regulating syntheses of nitric oxide, polyamines, and proteins, functional AA stimulate placental growth and the transfer of nutrients from mother to embryo or fetus to promote conceptus survival, growth, and development.

The intrauterine milieu is a complex mixture of substances originating from serum and endometrium that support blastocyst growth and development. The present study identified alterations in glucose and amino acids in response to an early rise in progesterone (P4), which accelerates blastocyst growth and development. Bred ewes received daily injections of either corn oil (CO) vehicle or P4 from 36 h postmating (Day 0) to either Day 9 or Day 12. Another group of ewes received P4 to Day 8 and the antiprogestin mifepristone (RU486) from Day 8 to Day 12. The total amount of glucose, aspartate (acidic amino acid), arginine and lysine (basic amino acids), and citrulline, asparagine, serine, glutamine, beta-alanine, and alanine (neutral amino acids) was greater in uterine flushings from early P4- than CO-treated ewes on Day 9. On Day 12, only arginine and lysine were higher in uterine flushings from P4-treated ewes, whereas citrulline was reduced. Glucose transporters, SLC2A1 and SLC5A1, were increased in uterine luminal (LE) and superficial glandular (sGE) epithelia of early P4-treated ewes on Days 9 and 12 but were reduced in endometria from ewes treated with both P4 and RU486 (P4+RU). SLC7A2B, a transporter of basic amino acids, increased in LE/sGE of P4- versus CO-treated ewes on Day 12 but was reduced in P4+RU-treated ewes. Thus, select nutrients are increased in the uterine lumen by P4 concomitant with the upregulation of epithelial transporters for glucose and basic amino acids, suggesting that these nutrients stimulate blastocyst growth and development.

Nutrients in uterine secretions are essential for development and survival of conceptuses (embryo and associated extraembryonic membranes) during pregnancy; however, little is known about changes in the amounts of specific nutrients in the uterine fluids of cyclic and pregnant ruminants. This study determined quantities of glucose, amino acids, glutathione, calcium, sodium, and potassium in uterine lumenal fluid from cyclic (Days 3-16) and pregnant (Days 10-16) ewes. Total recoverable glucose, Arg, Gln, Leu, Asp, Glu, Asn, His, beta-Ala, Tyr, Trp, Met, Val, Phe, Ile, Lys, Cys, Pro, glutathione, calcium, and sodium were greater in the uterine fluid of pregnant compared with cyclic ewes between Days 10 and 16. In cyclic ewes, only modest changes in the total amounts of glucose, Asn, Cit, Tyr, Trp, Met, Val, Cys, glutathione, calcium, and potassium were detected between Days 3 and 16. However, in pregnant ewes, amounts of glucose, Arg, Gln, Glu, Gly, Cys, Leu, Pro, glutathione, calcium, and potassium in uterine fluids increased 3- to 23-fold between Days 10 and 14 and remained high to Day 16. Of particular interest were increases in glucose, Arg, Leu, and Gln in uterine flushings of pregnant ewes between Days 10 and 16 of pregnancy. Total amounts of His, ornithine, Lys, Ser, Thr, Ile, Phe, Trp, Met, and Cit in uterine fluids also increased, but to a lesser extent during early pregnancy. These novel results indicate activation of pregnancy-associated mechanisms for transport of nutrients into the uterine lumen, and they provide a framework for future studies of nutrients, including glucose, amino acids, and glutathione, required to activate nutrient-sensing cell signaling pathways for growth, development, and survival of conceptuses, as well as for optimization of culture media for in vitro studies of conceptus development.

Nutrients, such as dietary lipids, can influence development of the central nervous system [1, 2]. Particularly, an increase in plasma concentrations of cholesterol, which is an essential constituent of all animal cells (especially of brain) and abundant in milk, is positively associated with enhancement of cerebrum weight gain and behavioral development [2–4]. Additionally, docosahexaenoic acid (DHA) is a component of complex lipid in membranes, nerve insulation, as well as a precursor for signaling molecules (including prostaglandins). The availability of long-chain polyunsaturated fatty acids, including DHA, also modulates eicosanoid metabolism and myelination during growth spurt of the brain [5,6]. Interestingly, dietary DHA supply enhances visual and neurological development both in prematurely born infants [7] and in term infants [8,9], while promoting problem-solving and childhood intelligence [10]. The beneficial influences of dietary cholesterol and DHA on brain development are currently interpreted as modification of the membrane structure and functions of membrane-associated proteins [11]. Nitric oxide (NO), synthesized from L-arginine by NO synthase (NOS), plays an important role in development and function of the brain [12]. All isoforms of the NOS [neuronal NOS (nNOS), endothelial NOS (eNOS), and inducible NOS (iNOS)] are present in neurological tissue. Of particular interest, nNOS activity is decreased when the protein is phosphorylated at the inhibition site of Ser 852, whereas eNOS activity is increased when the enzyme is phosphorylated at the activation site of Ser 1177 [12]. GTP-CH activity, Tetrahydrobiopterin (BH4), which is synthesized from GTP via the GTP cyclohydrolase I (GTP-CH) pathway [13], is an essential cofactor of NOS [14]. NO is a potent signal that regulates many physiological processes affecting behavior and cognitive function, including synaptic plasticity during long term potentiation and depression [15]. NO also acts as a retrograde messenger to stabilize or refine synapse during development, as a regulatory second messenger involved in the control of brain blood flow, and as an agent that contributes to both brain degeneration and neuropotection [16]. Additionally, NO promotes angiogenesis, redox state, cell immunity, and neuronal survival [14,17]. Cholesterol and DHA are known to regulate NO synthesis in vascular endothelial cells [17]. We hypothesized that dietary supplementation with cholesterol and DHA can influence the arginine-NO pathway in the brain of neonatal animals. This hypothesis was tested with piglets by measuring NOS activity, total and phosphorylated levels for the three NOS isoforms, as well as concentrations of arginine, BH4 and NADPH in brain. To determine whether the effects of DHA and cholesterol are specific to the neuronal tissue, we also analyzed the above parameters in skeletal muscle and liver.

Cholesterol and docosahexaenoic acid (DHA) are important nutrients for neural development of infants. However, little is known about the effect of cholesterol or DHA on concentrations of amino acids (AA) in neonatal tissues. This study was conducted with the piglet (an established model for studying human infant nutrition) to test the hypothesis that dietary supplementation with the lipids may modulate AA availability in tissues. Sixteen newborn pigs were nursed by sows for 24 h and then assigned to one of four treatment groups, representing supplementation with 0.0% (control), 0.2% cholesterol, 0.2% DHA, or cholesterol plus DHA to the basal milk-formula. All piglets were euthanized at 49 days of age. In brain, cholesterol supplementation reduced (P < 0.05) concentrations of glutamate, serine, glutamine, threonine, β-alanine, alanine, methionine, isoleucine, leucine, and γ-aminobutyrate but increased (P < 0.05) concentrations of glycine and lysine, whereas DHA supplementation similarly affected (P < 0.05) concentrations of the same AA (except for isoleucine and lysine) and taurine. In addition, concentrations of most AA in liver, muscle and plasma were substantially altered by dietary supplementation of cholesterol and DHA in a tissue-dependent manner. Further, DHA reduced concentrations of carnosine in skeletal muscle, as well as ammonia in both plasma and brain. The results reveal that cholesterol and DHA can regulate AA metabolism and availability in various tissues of piglets. These novel findings have important implications for designing the next generation of infant formula to optimize neonatal growth and development.

Dietary supplementation with a high level of zinc oxide (ZnO) has been shown to reduce the incidence of diarrhea in weanling pigs, but the underlying mechanisms remain largely unknown. Intestinal-mucosal mast cells, whose maturation and proliferation is under the control of the stem cell factor (SCF), play an important role in the etiology of diarrhea by releasing histamine. The present study was conducted to test the novel hypothesis that supplementing ZnO to the diet for weanling piglets may inhibit SCF expression in the small intestine, thereby reducing the number of mast cells, histamine release, and diarrhea. In Experiment 1, 32 piglets (28 days of age) were weaned and fed diets containing 100 or 3000 mg zinc/kg (as ZnO) for 10 days (16 piglets per group). In Experiment 2, two groups of 28-day-old piglets (8 piglets per group) were fed the 100- or 3000-mg zinc/kg diet as in Experiment 1, except that they were pair-fed the same amounts of feed. Supplementation with a high level of ZnO reduced the incidence of diarrhea in weanling piglets. Dietary Zn supplementation reduced expression of the SCF gene at both mRNA and protein levels, the number of mast cells in the mucosa and submucosa of the small intestine and histamine release from mucosal mast cells. Collectively, our results indicate that dietary supplementation with ZnO inhibits SCF expression in the small intestine, leading to reductions in the number of mast cells and histamine release. These findings may have important implications for the prevention of weaning-associated diarrhea in piglets.

These studies were conducted to determine whether ghrelin, a 28-amino acid peptide produced mainly by the stomach, was involved in tryptophan-mediated appetite stimulation in swine. In experiment 1, 36 crossbred (Long WhitexLarge White) barrows were used in a 2x3 factorial design to determine the effects of food intake (ad libitum versus limit fed) and tryptophan level (0.12%, 0.19% and 0.26%) on growth performance as well as ghrelin expression, plasma insulin, ghrelin and leptin levels. Ad libitum fed pigs gained more weight, but had poorer feed conversion than limit fed pigs. Weight gain, food intake and feed conversion all improved with increased ingestion of dietary tryptophan. Ad libitum feeding increased plasma insulin. Plasma insulin was unaffected by the level of dietary tryptophan. However, plasma leptin was significantly lower in pigs fed 0.19% tryptophan compared to those fed 0.12% tryptophan. Plasma ghrelin levels and ghrelin mRNA level in gastric fundus and duodenun was significantly higher in pigs fed 0.19% and 0.26% tryptophan diet compared with pigs fed 0.12%. In the second experiment, 18 crossbred barrows were divided into three treatments involving oral infusion of saline, tryptophan (40mg/kg BW) or 5-hydroxytryptophan (40mg/kg BW). Plasma ghrelin levels at 20, 40 and 60min after infusion of tryptophan were higher than after saline and 5-hydroxytryptophan infusion, 5-hydroxytryptophan infusion induced lower food intake than saline infusion, and tryptophan infusion increased food intake 2, 8 and 24h after infusion. In conclusion, oral tryptophan ingestion increased ghrelin expression in gastric fundus and plasma ghrelin level.

Sympathetic vasoconstriction is attenuated in exercising muscle by locally generated vasodilators, including NO. Skeletal muscle also produces reactive oxygen species (ROS), such as superoxide (O 2 − ), which inactivates NO. We, therefore, hypothesized that excessive ROS production would result in enhanced sympathetic vasoconstriction in exercising muscle. To increase O 2 − by activating NADPH oxidase, rats underwent chronic infusion of angiotensin II (Ang II) or unilateral renal artery stenosis (2K1C) to increase endogenous Ang II. At rest, sympathetic nerve stimulation (range: 1 to 5 Hz) evoked similar graded decreases in femoral vascular conductance (range, −34% to −66%) in rats infused with vehicle, Ang II, or norepinephrine and in 2K1C or sham-operated rats. These sympathetically mediated decreases in femoral vascular conductance were markedly attenuated during hindlimb contraction in the vehicle, norepinephrine, and sham rats (range, −3% to −26%) and to a lesser degree in the Ang II (range, −16% to −47%) and 2K1C (range, −16% to −45%) rats. In muscles from Ang II and 2K1C rats, ROS were elevated and the NADPH oxidase subunit gp91 phox was upregulated. The O 2 − scavenger tempol restored the normal attenuation of sympathetic vasoconstriction in the contracting hindlimbs of the Ang II and 2K1C rats, but this effect was prevented by pretreatment with an NO synthase inhibitor. Taken together, these data indicate that chronically elevated Ang II increases muscle ROS, which disrupts the normal NO-dependent attenuation of sympathetic vasoconstriction. These findings may have implications for muscle oxidative stress and sympathetic vasoregulation when the renin–angiotensin system is chronically activated.

This study was conducted to examine the relationship between soya-bean agglutinin and cholecystokinin in stimulating pancreatic enzyme synthesis in rats. In experiment 1, 30 rats were given daily gastric infusions of 0, 3·5, 7·0, 10·5, or 14·0 mg of soya-bean agglutinin (no.=6) for 14 days. Compared with the control group, soya-bean agglutinin reduced weight gains, enhanced absolute and relative dry pancreatic weights, elevated serum cholecystokinin levels, and stimulated cholecystokinin mRNA expression in the intestine (PP

This study was conducted to investigate the mechanism for the effect of elevated levels of dietary zinc oxide (ZnO) in enhancing the intestinal growth of weanling piglets. In Experiment 1, 4-wk-old (8.1 +/- 0.6 kg) crossbred barrows (n = 36) were assigned randomly to 1 of the 2 dietary groups, with 6 pens/group (3 pigs/pen). One group was fed the basal diet containing 100 mg Zn/kg diet. The other group was fed the basal diet supplemented with ZnO to provide 3000 mg Zn/kg diet. Pigs consumed their feed ad libitum for 14 d. In Experiment 2, 4-wk-old (7.6 +/- 0.16 kg) crossbred barrows (n = 16) were housed individually and assigned to 1 of the 2 dietary groups (8 pigs/group) as in Experiment 1, except that the 2 groups were pair-fed the same amount of feed. At the end of a 14-d treatment period, all of the pigs in both Experiments 1 and 2 were weighed, feed consumption was measured, and blood samples were collected for assays of insulin-like growth factor-I (IGF-I). In addition, 1 pig from each pen in Experiments 1 and 2 was selected randomly to obtain the small-intestinal mucosa for analyzing IGF-I and IGF-I receptor (IGF-IR) gene expression and to determine the small-intestinal morphology. In Experiment 1, dietary supplementation of ZnO increased (P < 0.05) the daily body weight gain and daily feed intake. In Experiment 2, dietary supplementation of ZnO increased (P < 0.05) the daily body weight gain and feed conversion efficiency. In both experiments, the villous height of the small-intestinal mucosa and both the mRNA and protein levels for IGF-I and IGF-IR in the small intestine were markedly enhanced (P < 0.05) by feeding elevated levels of Zn. Serum IGF-I levels did not differ between the control and Zn-supplemental groups in either experiment. Collectively, these results suggest that dietary Zn supplementation exerts its beneficial effects on the intestinal growth of weanling piglets through increasing IGF-I and IGF-IR expression in the small-intestinal mucosa.

Insufficient placental growth is a major factor contributing to intrauterine growth retardation in mammals. There is growing evidence that putrescine produced from arginine (Arg) and proline via ornithine decarboxylase is a key regulator of angiogenesis, embryogenesis, as well as placental and fetal growth. However, the underlying mechanisms are largely unknown. The present study tested the hypothesis that putrescine stimulates protein synthesis by activating the mechanistic target of rapamycin (mTOR) signaling pathway in porcine trophectoderm cell line 2 cells. The cells were cultured for 2 to 4 days in customized Arg-free Dulbecco modified Eagle Ham medium containing 0, 10, 25, or 50 μM putrescine or 100 μM Arg. Cell proliferation, protein synthesis, and degradation, as well as the abundance of total and phosphorylated mTOR, ribosomal protein S6 kinase 1, and eukaryotic initiation factor 4E-binding protein-1 (4EBP1), were determined. Our results indicate that putrescine promotes cell proliferation and protein synthesis in a dose- and time-dependent manner, which was inhibited by difluoro-methylornithine (an inhibitor of ornithine decarboxylase). Moreover, supplementation of culture medium with putrescine increased the abundance of phosphorylated mTOR and its downstream targets, 4EBP1 and p70 S6K1 proteins. Collectively, these findings reveal a novel and important role for putrescine in regulating the mTOR signaling pathway in porcine placental cells. We suggest that dietary supplementation with or intravenous administration of putrescine may provide a new and effective strategy to improve survival and growth of embryos/fetuses in mammals.

Chronic viral infections lead to persistent immune activation, which is alleviated by eradicating or suppressing the infection. To understand the effects of interferon treatment on immune system activation by chronic infections, we evaluated kinetic patterns of a broad spectrum of serum biomarkers during HCV treatment in HIV/HCV co-infected patients. HCV viral load and 50 biomarkers were analysed at baseline and 27 time points during pegylated interferon-alpha and ribavirin (IFN/RBV) treatment of 12 HIV/HCV co-infected patients. We evaluated biomarker changes from baseline for each time point and biomarker correlations with clinical parameters, treatment response and liver histopathology. IL-1α, IL-12p40, IL-1RA, IP-10, MIG, MIP-1α/1β, HGF, sCD40L, TRAIL and leptin increased in the first day. IL-12p70, IL-17A, IL-10, GROα, IL-8, MCP-3, IL-4 and M-CSF peaked later during week 1. IL-1α, HGF, IP-10, MIP-1α, TRAIL, sCD40L, IL-10, IL-12p70, MCP-3, FGFb, ENA-78, TGF-β, IL-2, IFN-γ, IL-6, IL-15, IL-7 and PDGF-BB decreased below baseline over the course of treatment. Higher BMI, baseline HCV viral load and leptin levels were associated with lack of sustained virologic response. ENA-78 was associated with sustained viral response. Positive correlations were found between liver inflammation and baseline CD4 count, sVCAM and HGF; fibrosis stage and HGF; liver steatosis, BMI and leptin. Our findings suggest IFN/RBV treatment initially increases levels of several biomarkers, but eventually leads to a decline in many immune markers. These findings shed light on the relationship between IFN treatment and immune activation by chronic viral infections, such as HCV.

Embryonic loss and intrauterine growth restriction (IUGR) are significant problems in humans and other animals. Results from studies involving pigs and sheep have indicated that limited uterine capacity and placental insufficiency are major factors contributing to suboptimal reproduction in mammals. Our discovery of the unusual abundance of the arginine family of amino acids in porcine and ovine allantoic fluids during early gestation led to the novel hypothesis that arginine plays an important role in conceptus (embryo and extra-embryonic membranes) development. Arginine is metabolized to ornithine, proline, and nitric oxide, with each having important physiological functions. Nitric oxide is a vasodilator and angiogenic factor, whereas ornithine and proline are substrates for uterine and placental synthesis of polyamines that are key regulators of gene expression, protein synthesis, and angiogenesis. Additionally, arginine activates the mechanistic (mammalian) target of rapamycin cell signaling pathway to stimulate protein synthesis in the placenta, uterus, and fetus. Thus, dietary supplementation with 0.83 % L-arginine to gilts consuming 2 kg of a typical gestation diet between either days 14 and 28 or between days 30 and 114 of pregnancy increases the number of live-born piglets and litter birth weight. Similar results have been reported for gestating rats and ewes. In sheep, arginine also stimulates development of fetal brown adipose tissue. Furthermore, oral administration of arginine to women with IUGR has been reported to enhance fetal growth. Collectively, enhancement of uterine as well as placental growth and function through dietary arginine supplementation provides an effective solution to improving embryonic and fetal survival and growth.

Importance An increasing number of patients with type 2 diabetes are treated with high doses of insulin. Such treatment is associated with weight gain, hypoglycemia, and high treatment burden. Objective To assess the effectiveness and safety of adding a glucagon-like peptide 1 receptor agonist to the treatment regimen of patients with type 2 diabetes requiring therapy with high-dose insulin. Design, Setting, and Participants This clinical trial was a double-blind, placebo-controlled, randomized (1:1) study with 6 months of follow-up, conducted from August 13, 2012, to February 9, 2015, at ambulatory clinics at the University of Texas Southwestern Medical Center and Parkland Hospital. Participants were 71 patients with uncontrolled type 2 diabetes (glycated hemoglobin level, 7.5%-11.0%) using more than 1.5 U/kg/d of insulin. Interventions Subcutaneous injection of liraglutide (1.8 mg/d) or matching placebo for 6 months. Main Outcomes and Measures The primary outcome was the change in glycated hemoglobin level. Secondary outcomes were changes in weight, hypoglycemia rate, insulin dosage, and quality-of-life measures. Results Among 71 patients, 45 (63%) were female. The mean (SD) age of patients was 54.2 (7.4) years, with a mean (SD) type 2 diabetes duration of 17.9 (8.4) years and a mean (SD) total daily dose of insulin of 247.0 (95.1) U. Ninety-three percent (66 of 71) of participants completed all scheduled visits. The glycated hemoglobin level improved from a mean (SD) of 9.0% (1.2%) to 7.9% (1.1%) in the liraglutide group ( P P = .002). Weight decreased from a mean (SD) of 114.6 (21.4) kg to 113.6 (20.8) kg in the liraglutide group vs a mean (SD) increase from 116.1 (26.6) kg to 117.2 (27.2) kg in the placebo group, with a treatment difference of −2.3 kg (95% CI, −4.3 to −0.4 kg) ( P = .02). The total daily dose of insulin decreased 11.5% (95% CI, −21.8% to −1.1%) in the liraglutide group ( P = .20). The hypoglycemia rate was higher in the first month after initiation of liraglutide compared with placebo (2.30 vs 0.91 events per person-month, P = .01), while the overall hypoglycemia rate over the entire follow-up was similar between groups ( P = .11). Glycemia control perception, satisfaction with insulin treatment, and willingness to continue insulin use improved more in the liraglutide group. Conclusions and Relevance Liraglutide added to high-dose insulin therapy improved glycemic control, decreased body weight, and enhanced treatment satisfaction in this difficult-to-treat patient population with high-dose insulin requirements. Further studies are warranted to confirm these findings and evaluate the long-term risk and benefit of this treatment option. Trial Registration clinicaltrials.gov Identifier:NCT01505673

Branched-chain amino acids (BCAA) are actively taken up and catabolized by the mammary gland during lactation for syntheses of glutamate, glutamine and aspartate. Available evidence shows that the onset of lactation is associated with increases in circulating levels of cortisol, prolactin and glucagon, but decreases in insulin and growth hormone. This study determined the effects of physiological concentrations of these hormones on the catabolism of leucine (a representative BCAA) in bovine mammary epithelial cells. Cells were incubated at 37 °C for 2 h in Krebs buffer containing 3 mM d-glucose, 0.5 mM l-leucine, l-[1-14C]leucine or l-[U-14C]leucine, and 0–50 μU/mL insulin, 0–20 ng/mL growth hormone 0–200 ng/mL prolactin, 0–150 nM cortisol or 0–300 pg/mL glucagon. Increasing extracellular concentrations of insulin did not affect leucine transamination or oxidative decarboxylation, but decreased the rate of oxidation of leucine carbons 2–6. Elevated levels of growth hormone dose dependently inhibited leucine catabolism, α-ketoisocaproate (KIC) production and the syntheses of glutamate plus glutamine. In contrast, cortisol and glucagon increased leucine transamination, leucine oxidative decarboxylation, KIC production, the oxidation of leucine 2–6 carbons and the syntheses of glutamate plus glutamine. Prolactin did not affect leucine catabolism in the cells. The changes in leucine degradation were consistent with alterations in abundances of BCAA transaminase and phosphorylated levels of branched-chain α-ketoacid dehydrogenase. Reductions in insulin and growth hormone but increases in cortisol and glucagon with lactation act in concert to stimulate BCAA catabolism for glutamate and glutamine syntheses. These coordinated changes in hormones may facilitate milk production in lactating mammals.

We recently reported that bacteria from the pig small intestine rapidly utilize and metabolize amino acids (AA). This study investigated the effect of L-arginine on the utilization of AA by pure bacterial strains (Streptococcus sp., Escherichia coli and Klebsiella sp.) and mixed bacterial cultures derived from the pig small intestine. Bacteria were incubated at 37°C for 3 h in anaerobic AA media containing 0-5 mmol/L of arginine to determine the effect of arginine on the bacterial utilization of AA. Amino acids in the medium plus cell extracts were analyzed by high-performance liquid chromatography. Results indicated concentration-dependent increases in the bacterial utilization of arginine and altered fluxes of arginine into ornithine and citrulline in the bacteria. Net glutamine utilization increased in pure bacterial strains with increased concentrations of arginine. With the addition of arginine, net utilization of threonine, glycine, phenylalanine and branched-chain AA increased (P

This study investigated the metabolism of select amino acids (AA) in bacterial strains (Streptococcus sp., Escherichia coli and Klebsiella sp.) and mixed bacterial cultures derived from the jejunum and ileum of pigs. Cells were incubated at 37°C for 3 h in anaerobic media containing 0.5–5 mM select AA plus [U-14C]-labeled tracers to determine their decarboxylation and incorporation into bacterial protein. Results showed that all types of bacteria rapidly utilized glutamine, lysine, arginine and threonine. However, rates of the utilization of AA by pure cultures of E. coli and Klebsiella sp. were greater than those for mixed bacterial cultures or Streptococcus sp. The oxidation of lysine, threonine and arginine accounted for 10% of their utilization in these pure bacterial cultures, but values were either higher or lower in mixed bacterial cultures depending on AA, bacterial species and the gut segment (e.g., 15% for lysine in jejunal and ileal mixed bacteria; 5.5 and 0.3% for threonine in jejunal mixed bacteria and ileal mixed bacteria, respectively; and 20% for arginine in ileal mixed bacteria). Percentages of AA used for bacterial protein synthesis were 50–70% for leucine, 25% for threonine, proline and methionine, 15% for lysine and arginine and 10% for glutamine. These results indicate diverse metabolism of AA in small-intestinal bacteria in a species- and gut compartment-dependent manner. This diversity may contribute to AA homeostasis in the gut. The findings have important implications for both animal and human nutrition, as well as their health and well-beings.

Dietary amino acids (AA) are crucial for animal growth, development, reproduction, lactation, and health. However, there is a scarcity of information regarding complete composition of “nutritionally nonessential AA” (NEAA; those AA which can be synthesized by animals) in diets. To provide a much-needed database, we quantified NEAA (including glutamate, glutamine, aspartate, and asparagine) in feed ingredients for comparison with “nutritionally essential AA” (EAA; those AA whose carbon skeletons cannot be formed by animals). Except for gelatin and feather meal, animal and plant ingredients contained high percentages of glutamate plus glutamine, branched-chain AA, and aspartate plus asparagine, which were 10–32, 15–25, and 8–14% of total protein, respectively. In particular, leucine and glutamine were most abundant in blood meal and casein (13% of total protein), respectively. Notably, gelatin, feather meal, fish meal, meat and bone meal, and poultry byproduct had high percentages of glycine, proline plus hydroxyproline, and arginine, which were 10–35, 9.6–35, and 7.2–7.9% of total protein, respectively. Among plant products, arginine was most abundant in peanut meal and cottonseed meal (14–16% of total protein), whereas corn and sorghum had low percentages of cysteine, lysine, methionine, and tryptophan (0.9–3% of total protein). Overall, feed ingredients of animal origin (except for gelatin) are excellent sources of NEAA and EAA for livestock, avian, and aquatic species, whereas gelatin provides highest amounts of arginine, glycine, and proline plus hydroxyproline. Because casein, corn, soybean, peanut, fish, and gelatin are consumed by children and adults, our findings also have important implications for human nutrition.

Proline plays important roles in protein synthesis and structure, metabolism (particularly the synthesis of arginine, polyamines, and glutamate via pyrroline-5-carboxylate), and nutrition, as well as wound healing, antioxidative reactions, and immune responses. On a per-gram basis, proline plus hydroxyproline are most abundant in collagen and milk proteins, and requirements of proline for whole-body protein synthesis are the greatest among all amino acids. Therefore, physiological needs for proline are particularly high during the life cycle. While most mammals (including humans and pigs) can synthesize proline from arginine and glutamine/glutamate, rates of endogenous synthesis are inadequate for neonates, birds, and fish. Thus, work with young pigs (a widely used animal model for studying infant nutrition) has shown that supplementing 0.0, 0.35, 0.7, 1.05, 1.4, and 2.1% proline to a proline-free chemically defined diet containing 0.48% arginine and 2% glutamate dose dependently improved daily growth rate and feed efficiency while reducing concentrations of urea in plasma. Additionally, maximal growth performance of chickens depended on at least 0.8% proline in the diet. Likewise, dietary supplementation with 0.07, 0.14, and 0.28% hydroxyproline (a metabolite of proline) to a plant protein-based diet enhanced weight gains of salmon. Based on its regulatory roles in cellular biochemistry, proline can be considered as a functional amino acid for mammalian, avian, and aquatic species. Further research is warranted to develop effective strategies of dietary supplementation with proline or hydroxyproline to benefit health, growth, and development of animals and humans.

In this study, we determined the effects of L-arginine supplementation during early pregnancy on embryonic/fetal survival and growth in gilts. Gilts were housed individually in pens and fed twice daily 1 kg of a corn- and soybean meal-based diet supplemented with 0.0, 0.4, or 0.8% L-arginine (wt:wt) between d 0 and 25 of gestation (10 gilts/treatment). The diets were made isonitrogenous by addition of appropriate amounts of L-alanine. At d 25 of gestation, gilts were fed L-alanine or L-arginine and hysterectomized 30 min later to obtain uteri and conceptuses (embryos and associated fetal membranes and fluids). Dietary supplementation with 0.4 or 0.8% L-arginine enhanced (P < 0.05) its concentrations in maternal plasma (64 and 98%, respectively) as well as the vascularity of chorionic and allantoic membranes, compared with the control group. Reproductive performance [numbers of corpora lutea (CL) and fetuses, placental and fetal weights, and embryonic mortality] did not differ between the 0.4% Arg and control groups. However, supplementation with 0.8% L-arginine decreased (P < 0.05) uterine weight (-20%), total number of fetuses (-24%), CL number (-17%), total fetal weight (-34%), total volume of allantoic and amniotic fluids (-34 to 42%), concentrations of progesterone in maternal plasma (-33%), as well as total amounts of progesterone (-35%), estrone (-40%), and estrone sulfate (-37%) in allantoic fluid, compared with the control group. These results indicate that dietary supplementation with 0.8% L-arginine between d 0 and 25 of gestation, while increasing placental vascularity, adversely affects the reproductive performance of gilts.