Effects of Tetrabasic Zinc Chloride on the Diarrhea Rate, Intestinal Morphology, Immune Indices and Microflora of Weaned Piglets.

This study was aimed to investigate the effects of different dietary zinc sources on the diarrhea rate, intestinal morphology, immune indexes and intestinal microbial composition of weaned piglets. A total of 240 weaned piglets (Duroc × Landrace × Yorkshire), at the age of 21 days, were randomly assigned to five dietary treatments for a four-week feeding trial to determine the effects of different amounts of tetrabasic zinc chloride (TBZC) supplementation on intestinal morphology, intestinal immune indices and intestinal microflora in weaned piglets, compared with the pharmacological dose of ZnO. The dietary treatments included a negative control (CON), (T1) ZnO (ZnO, 1500 mg/kg), (T2) tetrabasic zinc chloride (TBZC, 800 mg/kg), (T3) tetrabasic zinc chloride (TBZC, 1000 mg/kg), and (T4) tetrabasic zinc chloride (TBZC, 1200 mg/kg). Each treatment comprised six replicate pens, with eight pigs (four barrows and four gilts) per pen. Dietary TBZC of 1200 mg/kg improved the duodenum villus height, jejunum villus height and crypt depth of ileum, and increased the ratio of villus height to crypt depth of ileum (p < 0.05). The dietary supplementation of TBZC at a dosage of 1200 mg/kg has the potential to increase the levels of immunoglobulin G (IgG) and immunoglobulin A (IgA) in the duodenal mucosa. Furthermore, it shows a significant increase in the levels of immunoglobulin A (IgA) in the ileum. Compared with CON, TBZC significantly (p < 0.05) decreased pH values of stomach contents. It also increased the number of Firmicutes in intestinal contents. Compared with CON, the abundance of Firmicutes in jejunum contents of other treatments was significantly improved (p < 0.05), while the abundance of Proteobacteria in ileum contents of high-zinc treatments (T2 and T5) was decreased (p < 0.05). In conclusion, dietary TBZC of 1200 mg/kg improved the digestibility of crude protein in weaned piglets, altered the intestinal morphology of piglets, changed the intestinal microflora of piglets, reduced the diarrhea rate, and significantly improved the development of the small intestine of weaned piglets, and its regulation mechanism on intestinal tract needs further study. In summary, TBZC is likely to be an effective substitute source for the pharmacological dose of ZnO to control diarrhea in weaned piglets.

Effects of Dietary Valine Chelated Zinc Supplementation on Growth Performance, Antioxidant Capacity, Immunity, and Intestine Health in Weaned Piglets.

This study was conducted to investigate the effects of dietary valine chelated zinc (ZnVal) supplementation on growth performance, antioxidant capacity, immunity, and intestine health in weaned piglets. A total of 240 healthy 35-day-old weaned piglets (Duroc × Landrace × Yorkshire, average weight 10.70 ± 0.14 kg) were randomly divided into five groups with six replicate pens and eight piglets per pen. Dietary treatments were a corn-soybean meal basal diet supplemented with 0, 25, 50, 75, and 100 mg/kg ZnVal, respectively. The experiment lasted for 28 days. Results showed that average daily gain (ADG) was increased (P < 0.05) by ZnVal with 75-100 mg/kg supplementation on days 15-28 and with 50-100 mg/kg supplementation on days 1-28. Supplementation of 25-100 mg/kg ZnVal reduced (P < 0.01) the diarrhea rate of weaned piglets on days 1 to 14 and 1 to 28. Dietary supplementation with 25-100 mg/kg ZnVal increased (P < 0.05) copper/zinc-superoxide dismutase (Cu/Zn-SOD) and decreased malonaldehyde (MDA) activities in the serum on day 14 and 28. Supplementation of 25-100 mg/kg ZnVal increased (P < 0.05) glutathione peroxidase (GSH-Px) activity in serum on day 14. Additionally, the supplementation of 75 mg/kg ZnVal significantly increased the activity of superoxide dismutase (SOD) and Cu/Zn-SOD in the liver (P < 0.05). Furthermore, the supplementation of 25-100 mg/kg ZnVal significantly increased the total antioxidant capacity (T-AOC) in the liver (P < 0.05). Higher (P < 0.05) concentrations of IgG in the serum were measured from piglets supplemented with 75-100 mg/kg ZnVal on day 14 and dietary supplementation with 25-100 mg/kg ZnVal increased the level of immunoglobulin G (IgG) in serum on day 28 (P < 0.05). In addition, higher (P < 0.05) concentrations of immunoglobulin A (IgA) in the duodenum and ileum were measured from piglets supplemented with 75 mg/kg ZnVal and the supplementation of 25-100 mg/kg ZnVal also showed a higher (P < 0.05) concentration of immunoglobulin G (IgG) in duodenum. Supplementation of 50-100 mg/kg ZnVal increased the villus height and villus height/crypt depth of jejunum (P < 0.05). Moreover, dietary supplementation with 75-100 mg/kg ZnVal showed a higher (P < 0.05) concentration of zinc in the liver and supplementation of 50-100 mg/kg ZnVal increased (P < 0.05) the concentration of zinc in the heart, spleen, and kidney. In conclusion, the present research showed that supplementation of ZnVal improves growth performance by increasing antioxidant capacity and immunity and regulating intestinal morphology and the optimal inclusion level of ZnVal was 65~80 mg/kg.

4-Octyl itaconate restricts STING activation by blocking its palmitoylation.

The cyclic guanosine monophosphate adenosine monophosphate synthase (cGAS)-stimulator of interferon genes (STING) axis plays a vital role in defending foreign pathogens and maintaining immune homeostasis. While substantial advances have been made in understanding the metabolic changes that occur during macrophage activation, little is known about how these metabolic changes affect the cGAS-STING axis. In this study, we identify that 4-octyl itaconate (4-OI), a derivative of itaconate, inhibits the activation of cGAS-STING. Furthermore, we show that 4-OI inhibits cGAS-STING-related antiviral immune responses and autoimmune inflammation. However, we find that endogenous itaconate does not affect cGAS-STING activation, indicating that 4-OI and itaconate function differently. Mechanistically, we find that 4-OI directly alkylates STING at Cys91, blocking STING palmitoylation and oligomerization. The alkylation of STING by 4-OI represents another type of post-translational modifications (PTMs) of STING. Our findings reveal a mechanism by which cGAS-STING function is regulated through 4-OI alkylation and provide insights into the crosstalk between different kinds of PTMs.

Effects of Pelleting and Long-Term High-Temperature Stabilization on Vitamin Retention in Swine Feed.

The objective of this study was to study the effect of pelleting and long-term high-temperature stabilization on the retention of vitamin A, vitamin E, vitamin B2, and vitamin B6 in swine feed. Piglet diets (diet 1 and 3) were pelleted after conditioning at 83 °C for 120 s, and were high-temperature stabilized at 90 °C for 8.5 min after pelleting; the finishing pig diets (diet 2, 4, and 5) were pelleted after conditioning at 82 °C for 90 s, and were high-temperature stabilized at 85 °C for 9 min after pelleting; the samples were obtained before condition, after condition, after pelleting, and after cooling. The contents of vitamin A and vitamin E in diets 1−5 and vitamin B2, and vitamin B6 in diets 3−5 were detected. The results showed that: (1) the conditioning process had no significant effect on the retention of vitamin A, vitamin E, vitamin B2, and vitamin B6 in all experimental diets (p > 0.05); (2) the pelleting process and high-temperature stabilization process after pelleting had different degrees of influence on vitamins, among which the stabilization process had a more significant effect on the retention of vitamins. After pelleting and long-term high-temperature stabilization, the retention of vitamin A, vitamin E, and B2, and vitamin B6 were 68.8−77.3%, 56.9−90.1%, 63.8−70.3%, and 60.1−67.0%, respectively. In the process of pelleting and long-term high-temperature stabilization, the retention of vitamin A, vitamin E, vitamin B2, and vitamin B6 in the feed were significantly reduced (p < 0.05). Therefore, vitamin loss during high temperature and over a long period of time is worth considering, and vitamins must be over-supplemented.

Effects of α-glycerol monolaurate on intestinal morphology, nutrient digestibility, serum profiles, and gut microbiota in weaned piglets.

This experiment was conducted to investigate the effects of dietary supplementation of α-glycerol monolaurate (α-GML) on the growth performance, nutrient digestibility, serum profiles, intestinal morphology, and gut microbiota of weaned piglets. A total of 96 healthy 28-d-old (Duroc × Landrace × Yorkshire) weaned piglets with body weight of 8.34 ± 0.05 kg were randomly divided into 2 treatment groups with 6 replicate pens and 8 piglets per pen. The control group was fed a basal diet and the experimental group was fed the basal diet supplemented with 1,000 mg/kg α-GML. The experiment lasted for 28 d. Dietary supplementation with α-GML had no effect on average daily gain, average daily feed intake, or gain to feed ratio in piglets (P > 0.05); however, it reduced (P < 0.05) diarrhea rate of piglets on days 15 to 28. The apparent total tract digestibility of dry matter (DM), crude protein (CP), ether extract (EE), and gross energy (GE) on day 14, and DM, organic matter, CP, EE, and GE on day 28 increased (P < 0.05) with α-GML supplementation. Moreover, higher (P < 0.05) glutathione peroxidase activity and interleukin-10 (IL-10) concentration, and lower (P < 0.05) malondialdehyde and tumor necrosis factor-α concentrations were observed in piglets supplemented with α-GML compared with the control group on day 14. Compared with the control group, the villus height/crypt depth in the duodenum and villus height in the jejunum and ileum were significantly greater (P < 0.05) in the α-GML group. Dietary α-GML supplementation significantly increased (P < 0.05) the relative abundance of Firmicutes, while decreasing (P < 0.05) Bacteroidota and Campilobacterota in the cecal contents; significantly increased (P < 0.05) the relative proportion of Lactobacillus and Blautia species, reduced (P < 0.05) Eubacterium_rectale_ATCC_33656, Campylobacter, and uncultured_bacterium_Alloprevotella species. Thus, dietary α-GML supplementation at 1,000 mg/kg reduces diarrhea rate, improves intestinal morphology, nutrient digestibility, antioxidant capacity, and immune status, and ameliorates gut microbiota in weaned piglets.

Glycerol monolaurate (GML) is naturally present in breast milk as well as other natural sources such as coconut oil and is widely used as a food additive. Dietary α-GML is used in animal production due to its safe-guarding health and growth-promoting effects. In the present study, α-GML was evaluated for growth performance, blood parameters, and intestinal health in piglets. Dietary α-GML helped piglets digest dry matter, crude protein, ether extract, and gross energy in feed. The blood parameters and intestinal structure of piglets fed the diet containing 1,000 mg/kg α-GML were improved. In addition, α-GML supplementation promoted the colonization of beneficial bacteria and inhibited the number of harmful bacteria. In the current study, dietary α-GML was responsible for improving the health status, intestinal morphology, and digestion and absorption of nutrients of weaned piglets with less diarrhea.

Protocol for evaluation and validation of TLR8 antagonists in HEK-Blue cells via secreted embryonic alkaline phosphatase assay.

Toll-like receptor 8 (TLR8) is a pattern recognition receptor that senses RNA degradation products and initiates immune responses. TLR8 overactivation is associated with autoimmune diseases. Herein, we describe the evaluation and validation of TLR8 antagonists in HEK-Blue cells via secreted embryonic alkaline phosphatase (SEAP) assay, WST assay, ITC and immunoblotting. These assays can facilitate the development of TLR8 antagonists; this protocol can also be adapted to analyze agonists and antagonists for other TLRs. For complete details on the use and execution of this protocol, please refer to Hu et al. (2018).

Orthosteric-allosteric dual inhibitors of PfHT1 as selective antimalarial agents.

Artemisinin-resistant malaria parasites have emerged and have been spreading, posing a significant public health challenge. Antimalarial drugs with novel mechanisms of action are therefore urgently needed. In this report, we exploit a "selective starvation" strategy by inhibiting Plasmodium falciparum hexose transporter 1 (PfHT1), the sole hexose transporter in P. falciparum, over human glucose transporter 1 (hGLUT1), providing an alternative approach to fight against multidrug-resistant malaria parasites. The crystal structure of hGLUT3, which shares 80% sequence similarity with hGLUT1, was resolved in complex with C3361, a moderate PfHT1-specific inhibitor, at 2.3-Å resolution. Structural comparison between the present hGLUT3-C3361 and our previously reported PfHT1-C3361 confirmed the unique inhibitor binding-induced pocket in PfHT1. We then designed small molecules to simultaneously block the orthosteric and allosteric pockets of PfHT1. Through extensive structure-activity relationship studies, the TH-PF series was identified to selectively inhibit PfHT1 over hGLUT1 and potent against multiple strains of the blood-stage P. falciparum Our findings shed light on the next-generation chemotherapeutics with a paradigm-shifting structure-based design strategy to simultaneously target the orthosteric and allosteric sites of a transporter.

Solubility of Omeprazole Sulfide in Different Solvents at the Range of 280.35-319.65 K.

Solubility data were measured for omeprazole sulfide in ethanol, 95 mass-% ethanol, ethyl acetate, isopropanol, methanol, acetone, n-butanol and n-propanol in the temperature range from 280.35 to 319.65 K by employing the gravimetric method. The solubilities increase with temperature and they are in good agreement with the calculated solubility of the modified Apelblat equation and the λh equation. The experimental solubility and correlation equation in this work can be used as essential data and model in the purification process of omeprazole sulfide. The thermodynamic properties of the solution process, including the Gibbs energy, enthalpy, and entropy were calculated using the van't Hoff equation.

Immune responses and immune-related gene expression profile in orange-spotted grouper after immunization with Cryptocaryon irritans vaccine.

In order to elucidate the immune-protective mechanisms of inactivated Cryptocaryon irritans vaccine, different doses of C. irritans theronts were used to immunize orange-spotted grouper (Epinephelus coioides). We measured serum immobilization titer, blood leukocyte respiratory burst activity, serum alternative complement activity, and serum lysozyme activity weekly. In addition, the expression levels of immune-related genes such as interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α), major histocompatibility complexes I and II (MHC I and II), and transforming growth factor-ß1 (TGF-ß1) were determined in spleen and gills. The results showed that the immobilization titer, respiratory burst activity, and alternative complement activity of immunized fish were significantly increased, and the levels of the last two immune parameters in the high-dose vaccine group were significantly higher than in the low-dose vaccine group. Serum lysozyme activity in the high-dose vaccine group was significantly higher than in the PBS control group. Vaccination also regulated host immune-related gene expression. For example, at 2- and 3- weeks post immunization, IL-1ß expression in the high-dose vaccine group spleen was significantly increased. At 4-weeks post immunization, the fish were challenged with a lethal dose of parasite, and the survival rates of high-dose vaccine group, low-dose vaccine group, PBS control group, and adjuvant control group were 80%, 40%, 0%, and 10% respectively. These results demonstrate that inactivated C. irritans vaccination improves specific and nonspecific immune responses in fish, enhancing their anti-parasite ability. These effects are vaccine antigen dose-dependent.

[Vasorelaxational effects of procyanidins on rabbit aorta in vitro and decreasing arterial blood pressure in vivo].

OBJECTIVE: To study the vasodilation effect of the procyanidin (PC) extracted from grape seeds on rabbit thoracic aortic rings in vitro, decreasing blood pressure in vivo and the possible mechanism. METHOD: Rabbits aortic rings were isolated and were divided into six groups including removal of endothelium, integrity of endothelium, 1 x 10(-5) mol X L(-1) indomethacin (Indo), 1 x 10(-5) mol x L(-1) propranolol (Prop), 1 x 10(-4) mol x L(-1) N(omega)-nitro-L-arginine (L-NNA) and 1 x 10(-5) mol x L(-1) methylene blue (MB). Then the thoracic aortic rings were treated with PC with cumulative concentrations of 1.25, 2.5, 5.0 mg x L(-1) respectively and the changes of tension were recorded, and investigate the effect of 40 mg x L(-1) PC on the contraction of aortic smooth muscles, thoracic aortic rings were pre-treated with NA (1 x 10(-8) to approximately 1 x 10(-5) mol x L(-1)), KCl (6.3 to approximately 100 mmol x L(-1)) and CaCl2 (1 x 10(-5) to approximately 1 x 10(-5) mol x L(-1)) followed by treatment with PC. Then, rabbits common carotid artery was intubated and arterial blood pressure in vivo was recorded. PC with cumulative concentrations of 4.0, 8.0, 16, 32, 64, 84 mg x kg(-1) was injected into vein and the changes of arterial blood pressure were observed. RESULT: PC could relax isolated rabbit aorta and showedan obvious concentration-dependent relaxation (r = 0. 63, P < 0.001). The relaxant effect of PC was significantly reduced by removal of endothelium and by treatment with nitric oxide synthase inhibitor L-NNA, or guanylyl cyclase inhibitor MB. In addition PC could decrease the dose response curves of aortic rings to NA, KCl and CaCl2. PC has a significant concentration-dependent negative effect on arterial blood pressure in vivo (r = 0.92, P < 0.001). CONCLUSION: PC has a vasodilation effect not only in an endothelium-dependent, nitric oxide involved manner, but in inhibition of calcium release and blockage of potential-dependent calcium channels. PC could decrease the rabbit's arterial blood pressure significantly in vivo.